BRPI0807836A2 - "1-BENZENOSULFONYL-1H-INDOL DERIVATIVES AS CCR9 ACTIVITY INHIBITORS". - Google Patents

Abstract

1-Benzenesulfonyl-1H-indoles, processes for their production and their use as pharmaceuticals, e.g. in the treatment of disorders which are mediated by CCR9, such as e.g. inflammatory bowel disease.

Description

Invention Patent Report for "1-BENZENOSULFONYL-1H-INDOL DERIVATIVES AS CCR9 ACTIVITY INHIBITORS".

The present invention relates to 1-benzenesulfonyl-1H-indoles, more specifically to 1-benzenesulfonyl-1H-indoles as inhibitors of CCR9 activity,

CC 25 chemokine ligand (CCL25), originally described as thymus expressed chemokine (TECK), plays a crucial role in the return of T cells to the small intestine by signaling a10 through the CC 9 chemokine receptor (CCR9). CCL25 is constitutively expressed within the small intestine, especially in the epithelial crypts, being weak or nonexistent in the colon and other mucosal surfaces. CCR9 is the only known receptor for TECK / CCL25. CCR9 expression strongly correlates with the ability of peripheral T lymphocytes to return to the small intestine. Most intestinal intraepithelial lymphocytes (IEL) and lamina propria lymphocytes (LPL) are CCR9 +, while a much lower percentage of circulating T cells in the blood is CCR9 +. CCR9 + T cells found in peripheral blood almost exclusively have the intestinal return receptor α4β7,

Blocking CCR9 with TECK / CCL25 antibody significantly inhibits return of T lymphocytes to the small intestine. In addition, there is a rigid location of CCR9 + TECK / CCL25 and LPL in the small intestine rather than the large intestine, suggesting a distinctive mechanism of lymphocyte recruitment in different segments of the gastrointestinal tract.

Studies have also suggested a role of TECK / CCL25 in integrating

T-endothelium lymphocytes in inflamed intestinal mucosa. There is increased TECK / CCL25 expression and enhanced LPL adhesion to the small intestinal mucosa following TNFα stimulation. CCR9 or anti-TECK / CCL25 desensitization could attenuate lymphocyte recruitment 30 to the small intestine microvessels. Thus, targeted blockade of CCL25-CCR9 interactions can provide effective therapeutic treatment in immune-mediated diseases, for example intestinal disorders, such as autoimmune and inflammatory diseases or conditions. T lymphocyte (T-cell) infiltration into the small intestine and colon has been specifically linked to the pathogenesis of Celiac disease, food allergies, rheumatoid arthritis, inflammatory bowel disease (IBD) including 5 Crohn's disease and ulcerative colitis, for example. including ulcerative proctitis. Diseases that are also described to be CCR9 mediated, for example, include allergic diseases, psoriasis, atopic dermatitis, asthma, fibrotic disorders, disorders and diseases that originate or are transplanted, for example graft rejection, and cancer, such as leukemia. (acute lymphocytic leukemia 10), solid tumor, thymoma, thymic carcinoma.

Now compounds have been discovered that exhibit amazing activity as CCR9 inhibitors.

In one aspect, the present invention provides the use of a compound of the formula

v-15 where

R-1 is (C 1-12) alkyl, (C-M2) alkoxy, (C 3-12) cycloalkyl, optionally fused with heterocyclyl, (C 5-12) cycloalkenyl, optionally fused with heterocyclyl, (C 6-12) aryl, optionally fused with heterocyclyl, heterocyclyl, halogen, halo (C1-4) alkyl, preferably CF3, (C1-4) alkoxy, halo (C1-4) alkoxy, (C1-4) alkylthio,

wherein heterocyclyl is aliphatic or aromatic heterocyclyl comprising 3 to 12 ring members and 1 to 4 heteroatoms selected from N, O, S, optionally ringed with another ring system,

wherein cycloalkyl, cycloalkenyl, aryl or heterocyclyl are unsubstituted or substituted by one or more halogen, halo (C1-4) alkyl, halo (C1-4) alkoxy, (C1-4) alkyl, (C1-4) alkoxy or (C1-4 alkylthio,

R 2 and R 3 are different from each other and independently are hydrogen, hydroxycarbonyl, alkoxycarbonyl, for example (C 1) alkoxycarbonyl, cyano, aminocarbonyl, wherein amino is NH 2, once or twice substituted amino or a cyclic amine, halogen, halo ( C1-4 alkyl, such as CF3, (C1-4) alkyl, (C1-4) alkoxy, (C1-4) alkylthio in free form or in the form of a salt in the manufacture of a

treatment for disorders that are mediated by CCR9 activity, preferably inflammatory bowel disease.

Preferably in a compound of formula (I)

R 1 is (C 1-12) alkyl, for example tert-butyl, (C 1-12) alkoxy, (C 3-12) cycloalkyl, optionally fused with heterocyclyl, (C 5-12) cycloalkenyl, optionally fused with heterocyclyl, ( C6-18) aryl, optionally fused to heterocyclyl, heterocyclyl, halogen, e.g. chlorine, halo (C1-4) alkyl, preferably CF3, (C1-4) alkoxy, halo (C1-4) alkoxy, (C1- -4) alkylthio,

wherein heterocyclyl is aliphatic or aromatic heterocyclyl comprising 3 to 12 ring members and 1 to 4 heteroatoms selected from N, O, S, optionally ringed with another ring system, for example 4-morpholin-4-yl or 4-oxazol -5-ila,

wherein cycloalkyl, cycloalkenyl, aryl or heterocyclyl are unsubstituted or substituted by one or more halogen, halo (C1-4) alkyl, halo (C1.4) alkoxy, (C1-4) alkyl, (C1-4) alkoxy or (C1-4) alkylthio,

R 2 and R 3 are different from each other and independently are hydrogen, hydroxycarbonyl, alkoxycarbonyl, for example (C 1) alkoxycarbonyl, such as, for example, methoxycarbonyl, cyano, aminocarbonyl, where amino is NH 2, amino substituted once or twice or a cyclic amine, for example amino, methylamino, dimethylamino, isopropylamino, piperidin

1-yl, morpholin-4-yl, 4-methyl-piperazin-1-yl, halogen, halo (C1-4) alkyl, such as CF3, (C1-4) alkyl, (C1-4) alkoxy, (C1-4) alkylthio.

In a compound of formula (I) each single defined substituent may be a preferred substituent, for example independently of each other as defined substituent.

Preferably in a compound of the formula (I)> R 1 is tert-butyl, CF 3, chloro, an aliphatic 6-membered ring system having 2 heteroatoms selected from N, O, such as, for example, morpholin-4-yl, and R2 and R3 are as defined above.

Preferably in a compound of formula (I), R 2 is hydrogen, cyano, hydroxycarbonyl, aminocarbonyl, wherein amino is NH 2, once or twice substituted amino 5 or a cyclic amine, for example amino is NH 2, methylamino, dimethylamino, isopropylamino Hydroxycarbonyl-isopropylamino, piperidin-1-yl, 4-methyl-piperazin-1-yl, morpholin-4-yl, and R 1 and R 3 are as defined above.

Preferably in a compound of formula (I) R 3 is hydrogen, cyano, chloro, hydroxycarbonyl, methoxycarbonyl, aminocarbonyl, sulfonylaminocarbonyl, for example, CO-NH-SO 2 - (4-tert-butyl phenyl), and R 1 and R 2 are as defined above.

In another aspect, the present invention provides a compound of the

formula

. -15 where

R1 is branched (C4-12) alkyl, preferably tert-butyl, (Cw2) alkoxy, (C3-12) cycloalkyl, optionally fused with heterocyclyl, (C5-12) cycloalkenyl, optionally fused with heterocyclyl, (C6-12) aryl optionally fused with heterocyclyl, heterocyclyl, chloro or fluoro, halo (C1-4) alkyl, preferably CF3, (C1-4) alkoxy, halo (C1-4) alkoxy, (C1-4) alkylthio,

wherein heterocyclyl is aliphatic or aromatic heterocyclyl comprising 3 to 12 ring members and 1 to 4 heteroatoms selected from N, O, S, optionally ringed with another ring system,

wherein cycloalkyl, cycloalkenyl, aryl or heterocyclyl are unsubstituted or substituted by one or more halogen, halo (C1-4) alkyl, halo (C1-4) alkoxy, (C1-4) alkyl, (C1-4) alkoxy or (C1-4) alkylthio,

R2 and R3 are different from each other and independently are

(N) hydrogen, hydroxycarbonyl, alkoxycarbonyl, for example, (Cm) alkoxycarbonyl, cyano, aminocarbonyl, wherein amino is NH2, amino substituted once or twice or a cyclic amine, with the proviso that if R2 is 4-methylpiperazin-1 -ylcarbonyl, then R2 is at the 3-position of the ring system, chloro, halo (C1-4) alkyl, such as CF3, (C1-4 alkyl), (CM) alkoxy, (CM) alkylthio with the proviso that

selected from the group consisting of

- 1- (4-tert-Butyl-benzenesulfonyl) -5-chloro-1H-indol-2-acid

carboxylic,

- 1- (4-tert-butyl-benzenesulfonyl) -1 H -indole-6-carboxylic acid,

indole-7-carboxylic,

- 1- (4-tert-butyl-benzenesulfonyl) -1 H -indole-3-carboxylic acid,

- 5-chloro-1- (4-morpholin-4-yl) -benzenesulfonyl) -1H-indol-3- acid

carboxylic,

a compound of the formula

Cl

COOH

and a compound of the formula

Cl

are deleted.

10

In another aspect, the present invention provides a compound

15

- 1- (4-tert-Butyl-benzenesulfonyl) -1H acid methyl ester

20

- 1- (4-tert-Butyl-benzenesulfonyl) -5-chloro-1H-indol-3- acid

5-chloro-1- (4-oxazol-5-yl) -benzenesulfonyl) -1H-indole-3-carboxylic acid,

- 5-chloro-1- (4-trifluoromethyl-benzenesulfonyl) -1H-indole-3-acid

carboxylic,

- 5-chloro-1- (4-chloro-benzenesulfonyl) -1H-indole-3- acid

carboxylic,

- 5-chloro-1- (4-morpholin-4-yl-benzenesulfonyl) -1H-indol-2-acid

carboxylic,

- 1- (4-tert-butyl-benzenesulfonyl) -1 H -indole-2-carboxylic acid,

- 1- (4-Morpholin-4-yl-benzenesulfonyl) -1H-indol-6-carboxylic acid methyl ester,

- 1- (4-Morpholin-4-yl-benzenesulfonyl) -1H-indol-6- acid

carboxylic,

- 1- (4-tert-Butyl-benzenesulfonyl) -5-chloro-1H-indol-2-carboxylic acid amide,

- [1- (4-tert-Butyl-benzenesulfonyl) -5-chloro-1H-indol-3-yl] -morpholin-4-yl-methanone,

- 2 - {[1- (4-tert-butyl-benzenesulfonyl) -5-chloro-1H-indol-3-carbonyl] amino} -2-methyl-propionic acid,

- [1- (4-tert-Butyl-benzenesulfonyl) -5-chloro-1H-indol-3-yl] - (4-methylpiperazin-1-yl) -methanone,

- 1- (4-tert-Butyl-benzenesulfonyl) -5-chloro-1 H -indol-3-carboxylic acid methylamide,

- 1- (4-tert-Butyl-benzenesulfonyl) -5-chloro-1 H -indol-3-carboxylic acid dimethylamide,

- [1- (4-tert-Butyl-benzenesulfonyl) -5-chloro-1H-indol-3-yl] -piperidin

1-yl-methanone,

- 1- (4-tert-Butyl-benzenesulfonyl) -5-chloro-1H-indol-3-carboxylic acid amide,

1- (4-tert-Butyl-benzenesulfonyl) -5-chloro-1H-indol-3-carboxylic acid isopropylamide,

-1- (4-tert-butyl-benzenesulfonyl) -1H-indol-6-carbonitrile, -1- (4-tert-butyl-benzenesulfonyl) -5-chloro-1 H -indol-2-carbonitrile,

- 1- (4-tert-butyl-benzenesulfonyl) -1H-indol-3-carbonitrile,

- 1- (4-tert-butyl-benzenesulfonyl) -1H-indole-6-carboxylic acid amide and

- 4-tert-Butyl-N- [1- (4-tert-Butyl-benzenesulfonyl) -1H-indol-6-one

carbonyl] benzenesulfonamide,

for example compounds 1 to 27 as set forth in TABLE 1 in the examples.

Compounds provided by the present invention are hereinafter referred to as "compound (s) of (according to) the present invention". A compound of the present invention includes a compound in any form, for example in free form, as a salt, as a solvate and as a salt and solvate.

In another aspect the present invention provides a compound of the present invention in the form of a salt such as a hydrochloride.

Such salts preferably include pharmaceutically acceptable salts.

Although pharmaceutically unacceptable salts are included, for example, for preparation / isolation / purification purposes.

The present invention includes a compound of the present invention in any isomeric form and in any isomeric mixture. The present invention also includes tautomers of a compound provided by the present invention where tautomers may exist.

In another aspect the present invention provides a process for the production of a compound of formula (I) or (II) comprising the steps

The. react a compound of the formula

wherein R2 and R3 are as defined above with a compound of formula O

CI-S- (x) (IV)

ο

where R1 is as defined above,

and isolating a compound of formula (I) or (II) obtained from the reaction mixture, and optionally also reacting a compound of formula (I) or (II) obtained to obtain another compound of formula (I) or (II).

That reaction is an amine sulfonylation reaction, for example

for example by reacting an amine with a sulfonyl chloride. Such reactions are known and may be carried out when appropriate, for example by reacting a compound of formula (III) with a compound of formula (IV) in organic solvent, for example, a polar organic solvent, such as N 1 N 10 dimethylformamide, in the presence of a base, such as a hydride, for example sodium hydride, or a sodium or potassium salt of an alcohol, such as tert-butyl alcohol.

To obtain another compound of formula (I) or (II) a compound of formula (I) or (II) obtained can be converted to another compound of formula (I) or (II), for example also by reacting a compound of formula (I) or (II) obtained, for example, a compound of formula (I) or (II) obtained may be subjected to other reactions, as appropriate, for example, such as conventional; for example, if R 1 in a compound of formula (I) or (II) is halogen, for example bromine, such halogen group may be subjected to nucleophilic substitution, for example, by heterocyclyl, such as aliphatic heterocyclyl comprising a nitrogen atom. in the heterocyclic ring, such as morpholine, in organic solvent, for example an ether, such as dioxane, for example, in the presence of rac-BINAP and K3PO4, and treating a mixture thus obtained with Pd (dba) 2 under an atmosphere. inert, for example, under argon.

In an intermediate of formula (III) or formula (IV) (starting materials), functional groups, if present, may optionally be in protected form or in salt form if a salt-forming group is present. Protecting groups, optionally present, may be removed to an appropriate stage, for example according to, for example, analogously to a method as conventional. For example, if in a compound of the formula II R is carboxy, carboxy may be in protected form and the protecting group may be removed to an appropriate reaction stage, for example after reaction of a compound of formula (III) with 5%. a compound of formula (IV). For example, if R1 or R2 in a compound of formula (III) is alkoxycarbonyl, for example (C1-4 alkoxycarbonyl), the alkoxy group may be removed, for example, as appropriate, for example as conventional, for example by treating a compound of formula (I) or (II) wherein R1 or R2 is alkoxycarbonyl with LiOH in organic solvent, for example an ether such as dioxane to obtain a compound of formula (I) or (II) in wherein R1 or R2 is hydroxycarbonyl.

Alternatively, to obtain a compound of formula (I) or (II), a compound of formula (III) wherein R1 or R2 is hydrogen or a substituent other than hydroxycarbonyl or alkoxycarbonyl may be reacted with a compound of formula (IV). ), to obtain a compound of formula (I) or (II) wherein R1 or R2 is hydrogen or a different substituent than hydroxycarbonyl or alkoxycarbonyl, and also by reacting such compound obtained to obtain a compound of formula (I) or (II) wherein R1 or R2 is hydroxycarbonyl, for example by reacting such a compound obtained with n-BuLi and an excess of CO2, to obtain a compound of formula (I) or (II) wherein R1 or R2 is hydroxycarbonyl. Such a hydroxycarbonyl group, for example, may also be reacted to obtain a compound of formula (I) or (II) wherein R1 or R2 is alkoxycarbonyl, for example by an esterification reaction, as appropriate, for example, such as conventional.

Intermediates (starting materials) of formula (III) and formula

(IV) are known or may be prepared according to, for example, analogously to a method as conventional or as described herein. Any compound described herein, for example, a compound of the present invention and intermediates of formula (III) and (IV) may be prepared as appropriate, for example, as, for example, analogously to a method as conventional, for example, or as specified here.

The compounds of the present invention, for example, including a compound of formula (I) or (II), exhibit pharmacological activity and are therefore useful as pharmaceuticals. Compounds of the present invention exhibit dose-dependent inhibition in

- Scintillation proximity test (SPA TEST)

- Eu-GTP CONNECTION TEST

- Calcium Mobilization Test (FLIPR TEST)

for example under conditions as conventional, for example under conditions as described herein, for example in the nanomolar range of IC50 to low micromolar.

Activity in the treatment of inflammatory bowel disease is,

for example, determined in an SCID mouse model of inflammatory bowel disease.

CINTILATION PROXIMITY TEST (SPA)

SPA PRINCIPLE Chemokines mediate their actions through seven receptors

coupled to the G protein crossing the transmembrane (GPCR) in the target cells. Binding of Ligand to GPCRs stimulates GTP / GDP exchange in heterotrimeric G proteins, composed of α, β, γ subunits. The agonist-bound GPCR initiates the guanine nucleotide cycle by catalyzing the α-subunit GDP disassociation, allowing endogenous GTP binding, and dissociation of the βγ complex. Ga-GTP and ββγ subunits each can activate effectors such as adenylyl cyclase, phospholipase C and ion channels (see, for example, Neer EJ, Cell; 80: 249-57 (1995)). Ga-GTP is inactivated by an intrinsic GTPase activity that hydrolyzes GTP to GDP; subsequently the GDP-containing G protein is ready during the next activation cycle. This process can be monitored in vitro by measuring the binding of hydrolysis resistant GTP analogs such as 5'-0- (3- [35S] thiophosphate

35

([S] -GTPyS), to cell membranes containing the receptor of interest. A GTPyS scintillation proximity assay (SPA) is shown to be a useful functional assay for monitoring CCR9 activation by TECK.

SPA is a homogeneous and versatile assay technology for rapid and sensitive testing of a wide range of biological processes. The assay format requires no separation step and is accessible for automation. The receptor-bearing membranes are coupled by half of glycoprotein to fluorescent wheat germ agglutinin coated beads (Amersham Bioscience, No. RNPQ 0001). Once immobilized, the receptor is very close to the count so that if the agonist-bound GPCR initiates the guanine nucleotide cycle, [35SJGTPyS (Amersham Bioscience, No. SJ1308) binds to the membrane. The radioactive molecule will be kept in close proximity as the falling particles stimulate the scintillant within the bead to emit light, which is then detected by a PMT-based scintillation counter. Unbound radioligand is too far from the bill to transfer energy and therefore passes undetected.

CELLS AND CELL CULTURE

Human CCR9 receptor transfected 300-19 mouse pre-B-cells are cultured in suspension in cell culture flasks (100 ml cell suspension in 162 cm2 cell culture flask) at 37 ° C in an atmosphere containing 5% CO 2 in RPMI 1640 medium supplemented with penicillin (100 lU / ml), streptomycin (0.1 mg / ml), L-glutamine (to 4.5 mM final concentration), 10% FBS, 1 mM sodium pyruvate, 0.05 μΜ 2-mercaptoethanol, 1.5 pg / ml puromycin and 20 mM HEPES. Cells are usable for -12 passages for membrane preparation (ie CCR9 receptor density is acceptably high enough). CCR9 expression is monitored by FACS analysis using Alexa Fluor 647-conjugated mouse anti-human CCR9 antibody. CCR9 expression must be no less than 50% positive cells by FACS with respect to Alexa isotype control. Fluorine. As an approximation, a culture of 10x105 cells / ml can be divided using a 1: 30-1: 50 dilution and cell density starting after 2 to 3 days (~ 4 to 5 days for a centrifuge bottle culture ). Cells are harvested at a density of 8-10 x 105 cells / ml by centrifugation at 300 to 1000 g for 10 minutes. In general, cells are cultured and expanded to yield approximately 1 x 1010 cells. The combined cell pellet is washed once in cold (without calcium and magnesium) PBS, resuspended by pipetting in cold membrane buffer at approximately 2 x 10 8 cells / ml, frozen in dry ice, and stored at -80 ° C. .

MEMBRANE CAP

Membrane Buffer pH = 7.5 (1000 ml): 7.5 mM Tris, 12.5 mM MgCl2, 0.3 mM EDTA, 1 mM EGTA, 250 mM Sucrose, sterile filtrate and stored at + 4 ° C

TYPE BUFFER (50 mlV.

Membrane Buffer 45 ml + 10% glycerol

PREPARATION OF MEMBRANES

Pipette the cell suspension solution into sturdy tubes and mix each solution. Transfer the homogenates to centrifuge tubes and centrifuge 10 minutes at 1000 g. Harvest the supernatants. 15 Add 20 ml fresh membrane buffer to each pellet, transfer to the original robust tubes and homogenize and centrifuge again. Harvest the supernatants. Centrifuge the combined supernatants at 40,000 g for 30 minutes. Resuspend each pellet in 3 ml cold homogenization buffer with a Dounce homogenizer. Determine protein concentration in homogeneous suspension (BIO RAD assay, reference

BSA). Bradford Method (Microassay Procedure). As an approximate

10

data, 1 x 10 6 cell results in a membrane yield of 10 to 20 mg protein. Storage aliquots at -80 ° C.

OPTIMIZED COMPOUND TESTS AND SOLUTIONS

HEPES / BSA Buffer: 50 mM HEPES (pH 7.4), 50 pg / ml BSA 2.5 x Assay Buffer: 50 mM HEPES pH 7.4, 50 pg / ml BSA, 25 mM MgCl2, 25 μΜ GDP 250 mM NaCl, 375 pg / ml saponin

TECK: TECK dilutions are prepared with 0.1% BSA in PBS to yield 20-fold TECK solution for GTP binding assay. For compound testing, a concentration of 7.4 μΜ TECK is used to give a final concentration of 0.37 μΜ in the reaction. Compound Dilution: Test compounds are dissolved in DMSO at 100-fold higher final concentration in the assay. Serial dilutions of these concentrated compound solutions are made in DMSO which is diluted 5-fold in HEPES / BSA buffer to generate 20x concentrated compound solutions containing a 20% (v / v) DMSO concentration. The final DMSO concentration in the assays is 1% (v / v).

Membrane Dilution: Before use, the membranes (2.4 mg / ml raw material; CCR9-1 batch) are diluted in HEPES / BSA buffer to give 60 pg / ml. 50 μΙ of this membrane is added to each well. (3 pg / well final assay concentration for CCR9-1 membrane batch).

Final assay condition for compound testing: 50 mM HEPES pH 7.4, 50 pg / ml BSA, 100 mM NaCI, 10 mM MgCl2, 10 μΜ GDP, 150 pg / ml Saponin, 0.37 μΜ TECK and 3 μg / membrane well.

TEST PROTOCOL

The assay is performed by the zero time format, which involves the sequential addition of test samples, membrane, radioligand and beads as separate additions without any preincubation.

Briefly, the membranes are incubated in the presence of a20 agonist and composed with [35S] GTPyS and scintillation beads for 1 hour at room temperature in a vibrating mixer. Using a liquid handling robot, the following reagents are shipped in a 96-well White & Clear Isoplate (Wallac, # 1450-515) in the following sequence:

40 μΙ Assay Buffer (20 mM HEPES pH 7.5, 100 mM NaCI,

10 mM MgCl 2, 1 μ MGDP, 10 μg / ml Saponin, 50 μg / ml BSA).

10 μΙ Human TECK / CCL25 agonist, 25 μg / ml (R&D Systems, No. 334-TK-025)

10 μΙ sample in 50% DMSO 50 μΙ membranes, 60 μg / ml in assay buffer

50 μΙ [1 nM 35SJGTPyS1 in assay buffer,

40 μΙ bead suspension, 18.75 mg / ml assay buffer. After incubation plates were centrifuged for 5 minutes at 1000 xg and counted on the MicroBeta Counter (EG&G Wallac) in ParaLux SPA counting mode.

DATA ANALYSIS Data analysis is performed with Excel program package.

Fit 4.0 (Microsoft). To determine the quality of the experimental window of the assay, factor Z 'is calculated using only control data (baseline and stimulated values). For this assay, Z 'is estimated to be 0.73 which indicates a large separation band and overall excellent assay quality.

Eu-GTP BINDING TEST The Principle of Eu-GTP BINDING TEST

A time-resolved fluorometric method for measuring G protein activation using a non-hydrolyzable, non-hydrolyzable europium-labeled GTP analog, Eu-GTP.

MATERIALS

RPMI 1640 Medium With (Made of Powder, Gibco No. 074-01800) Liquid Penicillin / Streptomycin Solution (Gibco No. 15140-

122)

■ 20 FBS (certified, obtained from Gibco [# 16000] and then inactivated)

by heat)

Sodium Pyruvate (Gibco No. 11360-039)

Puromycin (used as selection marker; Sigma n0 P

8833)

Complete Protease Inhibitor (Roche No. 1697498)

Alexa Fluor 647 Conjugated Mouse Anti-Human CDFR9 Antibody (Pharmingen No. 557975)

Alexa Fluor 647 conjugated lgG2a isotype control (BD Pharmingen No. 557715)

TECK (aa24-150-his6, BMP Tool Protein Database No.

BTP04-005213, Aliquots of TECK Raw Material Solution (5 mg / ml; -350 μΜ)) stored at -80 ° C. BSA (Roche Diagnostics GmbH No. 775827)

Eu-GTP (Perkin-Elmer Life Science, Wallac, Turku, Finland; product code: AD0260), kit contains the following components:

Eu-GTP (1.65 nmol). The lyophilized Eu-GTP was reconstituted with distilled water to yield a 10 μΜ Eu-GTP concentration. Aliquots of the reconstituted Eu-GTP were stored at -20 ° C.

GDP (2.3 μιποΙ)

Freeze dried GDP is reconstituted with distilled water to yield a GDP concentration of 2 mm. Reconstituted GDP aliquots are stored at -20 ° C.

VICTOR2® V Multilabel Counter (Perkin-Elmer Life Sciences, Wallac, Turku, Finland)

MultiScreen Vacuum Manifold (Millipore No. MAVM 0960R) cells and cell culture To be performed as described herein under "CELLS AND CULTURE"

CELLS "in the" Proximity Scintillation Assay (SPA) "

Membrane Buffer and Homogenization Buffer

To be performed as described herein under "membrane buffer and homogenization buffer" in the "Scintillation Proximity Assay (SPA)"

PREPARATION OF MEMBRANES

To be performed as described herein under "membrane preparation" in the "Scintillation Proximity Assay (SPA)"

OPTIMIZED COMPOUND TESTS AND SOLUTIONS

To be performed as described herein under "Optimized Compound Testing Buffers and Solutions" in the "Scintillation Proximity Assay (SPA)"

For Eu-GTP: Dilute Eu-GTP raw material solution to 100 nM in HEPES / BSA buffer before use. GTP Wash Solution: 10 X GTP Wash Solution is diluted 1:10 with distilled water and cooled on ice. Eu-GTP CONNECTION TEST PROTOCOL FOR COMPOUND TESTING

The Eu-GTP binding assay is performed in a final volume of 100 μΙ on Acro-Well filter plates. Assay components are added to the wells in the following order:

Add 40 μΙ assay buffer (2.5 X) to each well (Wells B2 to G12). Add 5 μΙ TECK (7.4 μΜ) to the wells in columns 2-11 and the final TECK concentration in the assay is 0.37 μΜ. Add 5 μΙ of 0.1% BSA to column 12 wells serving as the basal control. Add 5 μΙ of 10 each concentration of compounds (20 times final concentration in 20% DMSO) in triplicate in columns 3-11 (ie 3 wells per concentration). Add 5 μΙ of 20% DMSO to wells in column 2 and 12 which are the stimulated and basal controls respectively. The final DMSO concentration in all wells is 1% (v / v). Add 50 μΙ membranes (3 pg / sample) 15 to all wells and mix briefly at 800 rpm on a microtiter plate shaker (MS1 Minishaker). The plate is incubated for 30-min with slow shaking at 300 rpm on an orbital plate shaker (MTS 2/4 digital microtiter shaker). Add 10 μΙ of 100 nM Eu-GTP per well to yield a final concentration of 10 nM. The plate is incubated for a further 20 min with slow shaking at 300 rpm on the orbital plate shaker. The reaction is terminated by vacuum filtration and the filter plate is washed twice by vacuum filtration with 300 μΙ of ice-cold GTP wash buffer per well. Eu-GTP retained on the filter is measured with a VICTOR2® V Multilabel Counter (340 nm excitation / 615 nm emission, 25 0.4 ms delay, 0.4 ms window) within 30 min after the wash step .

TABLE A (PLATE OUTLINE)

1 2 3 4 5 6 7 8 9 10 11 12

Control Control

The stimulatedConc. 1 cone. 2 Cone. 3 Cone. 4 Cone. 5 Cone. 6 Cone. 7 Cone. 8 Cone. 9 basal

B DMSO Comp 1 Comp 1 Comp 1 Comp 1 Comp 1 Comp 1 Comp 1 Comp 1 Comp 1 Comp1 DMSO C DMSO Comp 1 Comp 1 Comp 1 Comp 1 Comp 1 Comp 1 Comp 1 Comp 1 Comp 1 Comp 1 Comp 1 Comp 1 Comp 1 Comp 1 Comp 1 Comp 1 Comp 1 Comp 1 Comp 1 Comp1 Comp1 DMSO And DMSO Comp 2 Comp 2 Comp 2 Comp 2 Comp 2 Comp 2 Comp 2 Comp 2 Comp 2 Comp 2 DMSO F 2 Comp 2 Comp 2 Comp 2 Comp 2 Comp 2 Comp 2 Comp 2 Comp 2 DMSO G DMSO Comp 2 Comp 2 Comp 2 Comp 2 Comp 2 Comp 2 Comp 2 Comp 2 Comp 2 DMSO H DATA ANALYSIS

The current Eu-GTP binding assay caused by agonist stimulation (= a) is compared to basal binding (= b) and the final result is calculated as a percentage of basal binding [percent in basal = (a / bx 100 ) - 100].

Dose-response curves for the percent stimulation calculated above baseline binding for each test compound is adjusted using the Excel XLfit® complement program (ID Business Solutions, Guilford, Surrey, UK) for the 4-parameter logistic equation. (Model 205):

y = A + ((B-A) / (1 + ((C / x) d))) where x is concentration values, y percentage of stimulation above basal bond corresponding to x values.

The adjusted parameters are:

A: curve bottom plateau, B: curve top plateau, C:

x-value in the middle of the curve (ie between the top and bottom plateaus), D: slope factor (also known as the Hill coefficient).

The IC50 for the assay is defined as the intermediate point between TECK-containing solvent control and unstimulated solvent control.

The Z1 value is calculated using only control data (6 basal and 6 stimulated values) for each experiment. The Z 'varies between

0.56 and 0.79 in all assays.

CALCIUM MOBILIZATION TEST A) THE PRINCIPLE OF CALCIUM MOBILIZATION TEST

Chemokine receptors are receptors of seven pertussis toxin (PTX) sensitive Gai-coupled transmembranes. Several studies have demonstrated activation of various signaling pathways for most chemokines and in multiple cell types, including elevation of cytosolic intracellular calcium ([Ca2 +],) concentration. This process can be monitored in vitro by measuring ([Ca2 +]) levels by means of calcium sensitive fluorescent dyes using a fluorometric imaging plate reader (FLIPR). Mobilization of intracellular calcium in 4-MOLT cells, as measured using FLIPR technology, is shown to be a useful functional assay for monitoring TECK CCR9 activation.

B) CELLS AND CELL CULTURE

The MOLT-4 T-cell human leukemia strain was obtained from

from the American Type Culture Collection (ATCC, Manassas, VA). MOLT-4 cells are cultured in medium, which is RPMI-1640 supplemented with 10% FCS, 2 mM L-glutamine, 100 U / ml penicillin and 100 pg / ml streptomycin at 37 ° C with 5% CO2. Human serum albumin (HSA) is obtained from ZLB Behring (Vienna, Austria) as a 20% solution.

C) CALCIUM MOBILIZATION TEST PROTOCOL

The following solutions are prepared:

HPSS: 7.01g NaCl, 0.4g KCl, 0.2g MgSO4.7H2O, 4.76g HEPES, 2g Glucose. H2O (at 1 L)

Working buffer (WB): 600 ml HPSS + 0.9 ml 1 M CaCl2 + 12 ml 1 M HEPES.

% BSA / WB: 60 ml WB + 0.06 g Bovine Serum Albumin (BSA; Sigma A7906).

© Probenecid Raw Material Solution: 356 mg Probenecid + 2.5 ml 1 N NaOH + 2.5 ml WB.

«» Probenecid Buffer: 350 ml WB + 3,5 ml Probenecid Raw Material Solution.

• Fluo-4 Solution: 50 pg Fluo-4, AM + 0.025 ml DMSO + 0.025 ml Pluronic F-127 (Invitrogen / Molecular Probes No. P3000MP; provided as 20% in DMSO).

'Tincture solution: 105 ml medium + 1,05 ml Probenecida raw material solution + 2,1 ml 1 M HEPES + 0,21 ml Fluo-4 solution.

® TECK: prepared in 0.1% BSA / WB

MOLT-4 cells are harvested and loaded with Fluo-4 / acetoxymethyl ester (Fluo-4 / ΑΜ) according to the manufacturer's instructions (Invitrogen / Molecular Probes, Eugene, OR). Briefly, cells are incubated (1 x 10 7 cells per 3 ml) in tincture solution for 60 min at 37 ° C and 5% CO 2. Subsequently, cells are washed twice with Probenecea buffer and pipetted into 96-well assay plates (black, light-bottomed polystyrene plates; Corning Costar No. 3603) at 2 x 105 cells and 0.075 ml per well and then centrifuged. at 1200 revolutions for 5 minutes for 3 to 4 min to evenly distribute the cells at the bottom of the plates. The plates are incubated for 60 min in the dark at room temperature (RT) to allow for deesterification of intracellular AM esters. Test compounds are first dissolved in DMSO, and 0.006 ml of these DMSO feedstock solutions are diluted in 0.194 ml WB (± 10 HSA) prior to injection into the cell plates (0.025 ml / well). After a 30 min incubation in darkness at RT, intracellular Ca2 + mobilization is monitored after TECK injection (to give an effective near-maximum concentration of at least ECso) using a FLIPR instrument (Molecular Devices, Ismaning / Munich, Germany). Baseline readings are taken 15 (at 3.5 s intervals) for 25 s prior to TECK injection (0.025 ml / well) followed by 1 s intervals during 80 s after TECK injection. Fluorescence readings are performed using default settings, and all data is normalized using the formula:

D) CALCULATION

Calcium Response = [Fmax - Fmin] / Fmin

where Fmax represents the maximum fluorescence response and Fmin the minimum fluorescence, baseline. Dose response curves for calcium response data for each test compound are adjusted using the Excel XLfit® complement program (ID Business Solutions, Guilford, 25 Surrey, UK) for the 4-parameter logistic equation (Model 205 ) to determine IC50 values.

The compounds of the present invention show activity in assays as described herein and a compound of the present invention is prone to show therapeutic activity in the treatment of disorders that are mediated by CCR9 activity.

Disorders that are mediated by CCR9 activity and that are likely to be successfully treated with a CCR9 inhibitor, for example, include disorders in which CCR9 activity plays a causative or contributory role, such as disorders associated with CCR9 binding. CCRQ., CCL25, for example, CCRQ-mediated return-mediated disorders of leukocytes in a subject.

Disorders as used herein include diseases.

Disorders that are likely to be mediated by CCR9 activity, for example, include:

- disorders associated with inflammation

for example including inflammatory (chronic) disorders, disorders related to bronchial inflammation, for example including bronchitis, cervix, for example including cervicitis, conjunctiva, for example conjunctivitis, esophagus, for example esophagitis, cardiac muscle, for example eg, myocarditis, rectum, eg, proctitis, sclerotic, eg, scleritis, gum, involving bone, pulmonary inflammation (alveolitis), airways, eg, asthma, such as bronchial asthma, acute respiratory distress syndrome (ARDS) ), inflammatory skin disorders such as contact hypersensitivity, atopic dermatitis; fibrotic disease (eg pulmonary fibrosis), encephalitis, inflammatory osteolysis,

- disorders associated with immune system conditions,

-20 immune system, such as autoimmune disorders, for example, including

Graves disease, Hashimoto's disease (chronic thyroiditis), multiple sclerosis, rheumatoid arthritis, arthritis, gout, osteoarthritis, scleroderma, lupus syndromes, systemic lupus erythematosis, psoriasis, inflammatory bowel disease, including Crohn's disease colitis, for example ulcerative colitis; sepsis, septic shock, autoimmune hemolytic anemia (AHA), autoantibody-triggered urticaria, pemphigus, nephritis, glomerulonephritis, Goodpastur's syndrome, ankylosing spondylitis, Reiter's syndrome, polymyositis, dermatomyositis, cytokine-mediated toxicity, interleukin-2 toxicity, areata, uveitis, lichen planus, bullous pemphigoid 30, myasthenia gravis, type I diabetes mellitus, immune-mediated infertility such as premature ovarian insufficiency, polyglandular insufficiency, hypothyroidism, pemphigus vulgaris, lymphoid pemphigus, paraneoplastic pemphigus, and associated autoimmune hepatitis hepatitis B virus (HBV) and hepatitis C virus (HCV), Addison's disease, autoimmune skin diseases such as psoriasis, dermatitis herpetiformis, bullous epidermolysis, linear IgA bullous dermatosis, acquired bullous epidermolysis, chronic bullous disease 5 pernicious anemia, haemolytic anemia, vitiligo, polyglandular syndromes au toimmune type I, type II and type III, Autoimmune Hypoparathyroidism, Autoimmune Hypophysitis, Autoimmune Oophoritis, Gestational Pemphigoid, Healing Pemphigoid, Mixed Essential Cryoglobulinaemia, Immune Thrombocytopenic Purpura, Goodpasture Syndrome, Autoimmune Neoplasms 10 , stiff-man syndrome, encephalomyelitis, acute disseminated encephalomyelitis, GuillainBarre syndrome, cerebellar degeneration, retinopathy, primary biliary sclerosis, autoimmune hepatitis sclerosing cholangitis, reactive arthritis, polymyositis / dermatomyositis, mixed connective tissue disease15 Bechet's dome, polyarteritis nodosa allergic angitis and granulomatosis (Churg-Strauss disease), polyangiitis overlap syndrome (hypersensitivity) vasculitis, Wegener's granulomatosis, Kawasaki disease temporal arteritis, sarcoidosis, cryopathy, Celiac disease,

- disorders associated with cytokine mediated toxicity,

for example including interleukin-2 toxicity,

- bone disorders,

for example including osteoporosis, osteoarthritis,

- disorders associated with the brain and nerves,

- neurodegenerative disorders, for example, including central nervous system disorders as well as disorders of the peripheral nervous system, for example, CNS disorders including central nerve infections, brain injuries, cerebrovascular disorders and their consequences, Parkinson's disease, corticobasal degeneration, disease neuron disorder, dementia including ALS, multiple sclerosis, traumatic disorders,

including trauma and inflammatory consequences of trauma, traumatic brain injury, stroke, post-stroke, traumatic post-injury, small vessel cerebrovascular disease, eating disorders; other dementias, for example, including Alzheimer's disease, vascular dementia, Lewy body dementia, frontotemporal dementia and chromosome 17-related parquinsonism, frontotemporal dementias including Pick's disease, progressive nuclear paralysis, corticobasal degeneration, Huntington's disease, degeneration thalamic, Creutzfeld-Jakob dementia, HIV dementia, schizophrenia with dementia, Korsakoff psychosis, cognitive-related disorders such as moderate cognitive impairment, age-related memory impairment, age-related cognitive decline, vascular cognitive impairment, disorders of attention deficit, attention deficit hyperactivity disorder, and memory disorders in children with learning disabilities; conditions associated with the hypothalamic-pituitary-adrenal geometric axis,

- neuronal disorders, for example, including neuronal migration disorders, hypotonia (reduced muscle tone), muscle weakness,

seizures, developmental delay (physical or mental impairment), mental retardation, failure to thrive, eating difficulties, lymphoma, microcephaly, symptoms affecting the head and brain, motor dysfunction;

- disorders associated with the eye,

including uveoritinitis, vitreoretinopathy, corneal disease, iritis, iridocyclitis, cataracts, uveitis, diabetic retinopathy, retinitis pigmentosa, conjunctivitis, keratitis,

- disorders associated with the gastrointestinal tract

for example including colitis, inflammatory bowel disease,

Crohn's disease, ulcerative colitis, peptic ulceration, gastritis, oseophagitis,

- disorders associated with the heart and vascular conditions for example, including cardiovascular disorders, for example,

including heart failure, heart infarction, cardiac hypertrophy, cardiac arrest, for example, including all forms of heart pumping insufficiencies such as high and low acute and chronic right or left systolic or diastolic performance, regardless of underlying cause; myocardial infarction (M1), M1 prophylaxis (primary and secondary prevention), acute MI treatment, prevention of complications; heart disorders, proliferative vascular disorders, vasculitides, polyarteritis nodosa, inflammatory consequences of ischemia, 5 ischemic heart disease, myocardial infarction, stroke, peripheral vascular disease, pulmonary hypertension,

ischemic disorders, for example including myocardial ischemia, for example stable angina, unstable angina, angina pectoris, bronchitis; asymptomatic arrhythmias such as all forms of atrial and ventricular tachyarrhythmias, atrial tachycardia, atrial agitation, atrial fibrillation, atrioventricular reentrant tachycardia, pre-excitation syndrome, ventricular agitation, ventricular fibrillation, bradycardic forms of arrhythmias; arrhythmia, chronic obstructive pulmonary disease,

hypertension, such as systolic or diastolic high blood pressure, for example, essential and secondary hypertension, for example, including hypertensive vascular disorders, such as primary as well as all types of secondary, endocrine, neurogenic renal hypertension, and the like;

peripheral vascular disorders in which arterial and / or venous flow is reduced resulting in an imbalance between blood supply and tissue oxygen demand, for example including atherosclerosis, chronic peripheral arterial occlusive disease (PAOD), acute arterial thrombosis and embolism, inflammatory vascular disorders, Raynaud's phenomenon, and venous disorders; atherosclerosis, a disease in which the vessel wall is reshaped, for example, including accumulation of cells, smooth muscle cells, and inflammatory monocyte / macrophage cells within the vessel wall;

hypotension,

- liver and kidney disorders,

for example, including kidney disorders, kidney disorders, eg

eg acute renal failure, acute kidney disease, liver disorders, eg cirrhosis, hepatitis, liver failure, cholestasis, acute / chronic hepatitis, sclerosing cholangitis, primary biliary cirrhosis, acute / chronic interstitial / glomerulonephritis, granulomatous diseases,

- disorders associated with stomach or pancreas conditions

for example including stomach upset for example

gastric ulcer, gastrointestinal ulcer, pancreatic disorders, pancreatic fatigue,

- respiratory tract and lung disorders

including lung disorders, chronic lung disease 10, acute respiratory distress syndrome (ARDS), asthma, asthmatic bronchitis, bronchiectasis, diffuse interstitial lung disorders, pneumoconioses, fibrosing aveolitis, pulmonary fibrosis,

- disorders associated with skin and tissue conditions

conjunctive

including eczema, atopic dermatitis,

contact, psoriasis, acne, dermatomyositis, Sjòrgen's syndrome, Churg-Struass syndrome, sunburn, skin cancer, wound, urticaria, toxic epidermal necrolysis,

- disorders associated with allergic conditions,

• 20 for example, including late type hypersensitivity, conjunc-

allergicitis, drug allergies, rhinitis, allergic rhinitis, vasculitis, contact dermatitis;

- disorders associated with angiogenesis,

for example, including inefficient ability to recruit blood supply, disorders characterized by modified angiogenesis, tumor-associated angiogenesis,

- disorders associated with cancer and cell overproliferation,

for example, including premalignant conditions, hyperactive disorders

proliferatives, cancers whether primary or metastatic, cervical and metastatic cancer, cancer originating from uncontrolled cell proliferation, solid tumors such as those described in W002066019, including non-small cell lung cancer, cervical cancer; tumor growth, lymphoma, B-cell or T-cell lymphoma, benign tumors, benign nonproliferative disorders, renal carcinoma, esophageal cancer, stomach cancer, renal carcinoma, bladder cancer, breast cancer, colon cancer, lung cancer, melanoma nasopharyngeal cancer, osteocarcinoma, ovarian cancer, uterine cancer; prostate cancer, skin cancer, leukemia, tumor neovascularization, angiomas, myelodysplastic disorders, non-responsiveness to normal death inducing signals (immortalization), increased cell motility and invasion, genetic instability, dysregulated gene expression, cancer (neuro ) endocrine (carcinoid), blood cancer, lymphocytic leukemia, neuroblastoma; soft tissue cancer, metastasis prevention,

- disorders associated with diabetic conditions,

including diabetes (type I diabetes, type II diabetes), diabetic retinopathy, insulin-dependent diabetes, diabetes mellitus, gestational diabetes), insulin hyposecretion, obesity;

- disorders associated with endometriosis, testicular dysfunction

res,

- disorders associated with infectious disorders, for example with chronic infectious conditions,

For example, including bacterial disorders, otitis media, disease

Lyme disease, trioditis, viral disorders, parasitic disorders, fungal disorders, malaria, eg malaria anemia, sepsis, severe sepsis, septic shock, eg endotoxin-induced septic shock, exotoxin-induced toxic shock, infectious shock ( true septic), septic shock caused by Gram-negative bacteria, pelvic inflammatory disease, AIDS, enteritis, pneumonia; meningitis, encephalitis, lymphatic filarial infection,

- disorders associated with myasthenia gravis,

- disorders associated with nephritis,

including glomerulonephritis, interstitial nephritis, gra

Wegener's nullomatosis, fibrosis,

- pain-related disorders, for example CNS1-associated disorders such as multiple sclerosis, backbone injury, sciatica, unsuccessful back surgery syndrome, traumatic brain injury, epilepsy, Parkinson's disease, post-stroke, and back vascular lesions in the brain and dorsal spine (eg, infarction, hemorrhage, vascular malformation);

non-central neuropathic pain, for example, including that associated with post-mastectomy pain, phantom sensation, reflex sympathetic dystrophy (RSD), trigeminal neuralgiaradioculopathy, postoperative pain, HIV / AIDS-related pain, cancer pain, metabolic neuropathies (e.g., diabetic neuropathy, vasculitic neuropathy secondary to connective tissue disease), paraneoplastic polyneuropathy associated, for example, with lung carcinoma, or leukemia, or lymphoma, or prostate, colon, or stomach carcinoma, trigeminal neuralgia, cranial neuralgias , and postherpetic neuralgia;

pain associated with peripheral nerve damage, central pain (ie from

due to cerebral ischemia) and various chronic pains, ie, lumbago, back pain (low back pain), inflammatory and / or rheumatic pain;

headache pain (eg, migraine with aura, migraine without aura, and other migraine disorders), episodic and chronic tension-type headache, tension-like headache, cluster headache, and chronic paroximal hemicrania;

visceral pain such as pancreatitis, intestinal cystitis, dysmenorrhea, irritable bowel syndrome, Crohn's disease, biliary colic, ureteral colic, myocardial infarction and pelvic cavity pain syndromes, for example, vulvodynia, orchialgia, urethral syndrome 15 and protatodynia;

acute pain, for example, postoperative pain, and post trauma pain;

- disorders associated with rheumatic disorders, for example including arthritis, rheumatoid arthritis, osteoarthritis, psoriatic arthritis, crystalline arthropathies, gout, pseudogout, calcium pyrophosphate deposition disease, lupus syndromes, erythematous systemic lupus, sclerosis, multiple scleroderma, , arterosclerosis, arteriosclerosis, spondyloarthropathies, systemic sclerosis, reactive arthritis, Reiter1 syndrome, ankylosing spondylitis, polymyositis,

- disorders associated with sarcoidosis,

- disorders associated with transplantation,

for example including transplant rejection crisis and other disorders following transplantation, such as organ or tissue (xeno) transplant rejection, for example for the treatment of recipients of, for example, heart, lung, heart-lung combined , liver, kidney, pancreatic, skin, corneal transplants, host versus graft disease, such as following bone marrow transplantation, is10 chemical reperfusion injury,

- birth control (through ovulation inhibition).

Although ovulation inhibition is not a disorder, controlling

(by inhibiting ovulation) is also meant to be encompassed by the definition of "Disorders that are likely to be mediated by CCR9 activity" according to the present invention.

Disorders that are likely to be mediated by CCR9, for example, preferably include

- autoimmune disorders,

- inflammatory disorders,

- allergic disorders,

- disorders following transplantation,

- cancer;

more preferably autoimmune disorders, inflammatory disorders, disorders following transplantation;

such as

celiac disease, food allergy, rheumatoid arthritis, inflammatory bowel disease (IBD), Crohn's disease, ulcerative colitis, psoriasis, atopic dermatitis, asthma, fibrotic diseases, diseases following transplantation, GVH rejection, cancer, leukemia (acute lymphocytic leukemia) , solid tumors, carcinoids, thymoma, thymic carcinoma,

preferably IBD, such as Crohn's disease, ulcerative colitis, for example, including ulcerative proctitis. In another aspect, the present invention provides

- a compound of the present invention for use as a pharmaceutical,

the use of a compound of the present invention as a pharmaceutical

therapeutic,

- the use of a compound of the present invention for the manufacture of a medicament,

for example, for the treatment of CCR9 activity-mediated disorders;

for example, a compound of the present invention for treating it

CCR9 activity-mediated disorders, such as disorders associated with disruption of CCR9 binding to CCL25, such as CCR9-mediated return-mediated disorders of leukocytes in a subject; for the treatment of CCR9 activity-mediated disorders,

Compounds of formula (I) and (II) are 1-benzenesulfonyl-1H

indoles.

In another aspect, the present invention provides

- the use of 1-benzenesulfonyl-1H-indole as a pharmaceutical,

-the use of a 1-benzenesulfonyl-1H-indole for the manufacture of

a medicine,

for example, for the treatment of CCR9 activity-mediated disorders; for example as indicated above.

In another aspect, the present invention provides a compound of the present invention for the manufacture of a medicament for the treatment of inflammatory bowel disease.

For pharmaceutical use, one or more compounds of the present invention may be used, for example a combination of two or more compounds of the present invention, preferably a compound of the present invention is used.

A compound of the present invention may be used as a pharmaceutical in the form of a pharmaceutical composition. In another aspect, the present invention provides a pharmaceutical composition comprising a compound of the present invention in combination with at least one pharmaceutically acceptable excipient, for example, appropriate carrier and / or diluent, for example, including fillers, binders, disintegrants, conditioners, and the like. flow, lubricants, sugars or sweeteners, fragrances, preservatives, stabilizers, wetting and / or emulsifying agents, solubilizers, salts for regulating osmotic pressure and / or buffers.

In another aspect, the present invention provides a pharmaceutical composition of the present invention for the

use of treating disorders that are mediated by CCR9 activity,

the use of a pharmaceutical composition of the present invention to treat disorders that are mediated by CCR9 activity;

The use of a pharmaceutical composition comprising a compound of the present invention for the treatment of inflammatory disease.

of the gut.

the use of a pharmaceutical composition comprising a compound of the present invention to treat disorders that are mediated by CCR9 activity,

Treatment of disorders (diseases) as used herein includes pro

phylaxis (prevention).

For such treatment, the appropriate dosage, of course, will vary depending upon, for example, the chemical nature and pharmacokinetic data of a compound of the present invention used, the individual host, the mode of administration and the nature and severity of the conditions being treated. . However, in general, for satisfactory results in larger mammals, for example humans, an indicated daily dosage includes a range

- from about 0.0001 g to about 1.5 g, such as 0.001 g to 1.5

g,

- from about 0.001 mg / kg body weight to about 20 mg / kg body weight, such as 0.01 mg / kg body weight to 20 mg / kg body weight, of a compound of the present invention, by eg administered in divided doses up to four times a day.

A compound of the present invention may be administered to larger mammals, for example humans, by similar modes of administration that conventionally used with other mediators, for example low molecular weight inhibitors, of CCR9 activity.

In another aspect, the present invention provides a method of treating disorders that are mediated by CCR9 activity, for example, including disorders as specified above whose treatment comprises administering to a subject in need of such treatment a therapeutically effective amount of a compound. of the present invention; for example, in the form of a pharmaceutical composition.

In another aspect, the present invention provides:

- a compound of the present invention for the manufacture of a medicament,

- the use of a compound of the present invention for the manufacture of a medicament, for example a pharmaceutical composition,

for the treatment of disorders that are mediated by CCR9 activity.

In another aspect, the present invention provides a method of treating inflammatory bowel disease whose treatment comprises administering to a subject in need of such treatment a therapeutically effective amount of a compound of the present invention; for example, in the form of a pharmaceutical composition.

A compound of the present invention may be administered by any conventional route, e.g., enterally, for example, including nasal, buccal, rectal, oral administration; parenterally, for example, including intravenous, intraarterial, intramuscular, intracardiac, subcutaneous, intraosseous, transdermal (diffusion through intact skin), transmucosal (diffusion through a mucous membrane), inhalational administration; topically; for example, including epicutaneous, intranasal, intratracheal administration; intraperitoneal (infusion or injection into the peritoneal cavity); epidural (epidural) (epidural injection or infusion); intrathecal (injection or infusion into cerebrospinal fluid); intravitreal (administration through the eye); or by means of medical devices, for example for local release, for example stents, for example in the form of coated or uncoated tablets, capsules, solutions (injectables), infusion solutions, solid solutions, suspensions, dispersions. solid dispersions; for example, in the form of ampoules, jars, in the form of creams, gels, pastes, inhaler powder, foams, tinctures, lip sticks, drops, sprays, or as suppositories.

For topical use, for example, including administration to the eye, satisfactory results may be obtained with local administration of a concentration of 0.5 to 10%, such as 1 to 3% several times of active substance daily, for example 2 to 10%. 5 times a day.

A compound of the present invention may be administered as a pharmaceutically acceptable salt, for example an acid addition salt or metal salt; or in free form; optionally in the form of a solvate. A compound of the present invention in the form of a salt has the same order of activity as a compound of the present invention in free form; optionally in the form of a solvate.

A compound of the present invention may be used for any method or used as described herein alone or in combination with one or more at least one other second drug substance.

In another aspect, the present invention provides:

A combination of a compound of the present invention with at least one second drug substance;

A pharmaceutical combination comprising a compound

of the present invention in combination with at least one second drug substance;

A pharmaceutical composition comprising a compound of the present invention in combination with at least one second subset.

substance and one or more more pharmaceutically acceptable excipient (s));

A compound of the present invention in combination with at least one second drug substance, for example, in the form of a pharmaceutical combination or composition, for use in any method as defined herein, for example.

A combination, a pharmaceutical combination or a pharmaceutical composition comprising a compound of the present invention and at least one second drug substance for use as a pharmaceutical;

Use as a pharmacist of a compound of the present invention in combination with at least one second pharmaceutical substance

maca, for example, in the form of a pharmaceutical combination or composition;

Use of a compound of the present invention for the manufacture of a medicament for use in combination with a second drug substance; for example for any therapeutic treatment such as

indicated here;

A method of treating CCR9 activity-mediated disorders in a subject in need thereof, comprising co-administering, concomitantly or in sequence, a therapeutically effective amount of a compound of the present invention and at least one of the following:

A drug substance, for example, in the form of a combination or pharmaceutical composition;

A compound of the present invention in combination with at least one second drug substance, for example, in the form of a combination or pharmaceutical composition, for use in the preparation of a

medicine for use in disorders mediated by CCR9 activity.

Combinations include fixed combinations wherein a compound of the present invention and at least one second drug substance are in the same formulation; wherein a compound of the present invention and at least one second drug substance in formulations are

stops are provided in the same package, for example with co-administration instruction; and free combinations in which a compound of the present invention and at least a second drug substance are packaged separately, but instruction for concomitant or sequential administration is given.

In another aspect, the present invention provides:

A pharmaceutical package comprising a first drug substance which is a compound of the present invention and at least

a second drug substance, in addition to instructions for combined administration;

A pharmaceutical package comprising a compound of the present invention in addition to instructions for combined administration with

at least one second drug substance;

A pharmaceutical package comprising at least one second drug substance in addition to instructions for administration in combination with a compound of the present invention.

Combination treatment according to the present invention

may provide improvements compared to simple treatment.

In another aspect, the present invention provides:

A pharmaceutical combination comprising an amount of a compound of the present invention and an amount of a second drug substance, wherein the amounts are suitable for

produce a synergistic therapeutic effect;

A method of improving the therapeutic utility of the compound of the present invention comprising co-administering, for example, concomitantly or in sequence, a therapeutically effective amount of the compound of the present invention and a second drug substance.

- A method for improving the therapeutic utility of a

a second drug substance comprising co-administering, for example, concomitantly or in sequence, a therapeutically effective amount of compound of the present invention and a second drug substance.

A combination of the present invention and a second subs

Drug substance as a combination pair may be administered by any conventional route, for example as set forth above for a compound of the present invention. A second drug may be administered in dosages as appropriate, for example, in dosage ranges that are similar to those used for simple treatment, or, for example, in the case of synergy, even lower conventional dosage ranges.

Pharmaceutical compositions according to the present invention may be manufactured according, for example, analogously, to a method as conventional, for example mixing, granulating, coating, dissolving or lyophilizing processes. Unit dosage forms 10 may contain, for example, from about 0.1 mg to about 1500 mg, such as 1 mg to about 1000 mg.

Pharmaceutical compositions comprising a combination of the present invention and pharmaceutical compositions comprising a second drug as described herein may be provided as appropriate, for example according to, for example, analogously, with a method as conventional, or as described herein for a composition. of the present invention.

By the term "second drug substance" is meant a chemotherapeutic drug, especially any chemotherapeutic agent, other than an agent of the present invention.

For example, a second drug substance as used herein includes:

- other CCR9 inhibitors than a compound of the present invention, for example, including antibodies and molecular weight compounds.

low,

- anti-inflammatory and / or immunomodulatory drugs,

- antiallergic drugs

- anticancer drugs

- anesthetic drugs - antidiarrheal drugs.

For treatment of IBD the term "second drug substance" is meant to include an anti-inflammatory drug and / or an immunomodulator, for example, including a drug that is active in preventing or treating IBD and / or active in treating manifestations. of IBD, for example, symptoms of IBD, such as an anesthetic drug or an anti-diarrheal drug.

Anti-inflammatory and / or immunomodulatory drugs which are prone to be useful in combination with a compound of the present invention include for example

- mediators, for example inhibitors, of mTOR activity, including rapamycin of the formula

41

HO ,,

H3CO 39

35

36 CH3 I 3 O 32; 1 to 30

28 ^ OH

-The

Η3 °° "'

25

11

OCH

3 H3C1 18 20 17 22 CH3 19 21 and rapamycin derivatives, for example, including

40-0-alkyl rapamycin derivatives, such as 40-0-alkyl rapamycin derivatives

O-hydroxyalkyl rapamycin, such as 40-0- (2-hydroxy) ethyl rapamycin (everolimus),

32-deoxo-rapamycin derivatives and 32-hydroxyrapamycin derivatives, such as 32-deoxorapamycin,

16-0-substituted rapamycin derivatives such as 16-pent-2-ynyloxy-32-deoxorapamycin, 16-pent-2-ynyloxy-32 (S or R) -dihydro-rapamycin, 16-pent

2-ynyloxy-32 (S or R) -dihydro-40-0- (2-hydroxyethyl) -rapamycin,

rapamycin derivatives which are acylated to the oxygen group at position 40, for example 40- [3-hydroxy-2- (hydroxymethyl) -2-methylpropanoate] rapamycin (also known as CCI779), rapamycin derivatives which are substituted at the position 40 via heterocyclyl, for example 40-epi- (tetrazolyl) -rapamycin (also known as ABT578),

so-called rapalogues, for example, as described in W09802441, W00114387 and W00364383, such as AP23573, and

compounds described under the name TAFA-93, AP23464, AP23675, AP23841 and biolimus (e.g., biolimus A9).

mediators, for example inhibitors, of calcineurin, for example cyclosporin A, FK 506;

- ascomycin having immunosuppressive properties, for example and

for example, ABT-281, ASM981;

- corticosteroids; cyclophosphamide; azathioprene; leflunomide; mi

zoribine;

- mycophenolic acid or salt; for example sodium mycophenolate

mofetil;

15-deoxyispergualin or an immunosuppressive homologue, analog or derivative thereof;

mediators, for example inhibitors, of bcr-abl tyrosine kinase activity;

- mediators, for example inhibitors, of receptor activity

tyrosine kinase c-kit;

mediators, for example inhibitors, of tyrosine kinase PDGF receptor activity, for example Gleevec (imatinib);

- mediators, for example inhibitors, of p38 MAP activity

kinase,

- mediators, for example inhibitors, of VEGF tyrosine kinase receptor activity,

- mediators, for example inhibitors, of PKC activity, for example as described in W00238561 or W00382859, for example, the

compound of Example 56 or 70;

- mediators, for example inhibitors, of JAK3 kinase activity, for example Nbenzyl-3,4-dihydroxy-benzylidene-cyanoacetamide α-cyano- (3,4-dihydroxy) -] N-benzylcinamamide ( Tyrphostin AG 490), prodigiosin 25-C (PNU156804), [4- (4'-hydroxyphenyl) amino-6,7-dimethoxyquinazoline] (WHI-P131), [4- (3'-bromo-4'-hydroxylphenyl ) -amino-6,7-dimethoxyquinazoline] (WHI-P154), [4- (3,5'-dibromo-4'-hydroxyphenyl) -amino-6,7-dimethoxyquinoline] zoline] WHI-P97, KRX-211, 3 - {(3R, 4R) -4-methyl-3- [methyl- (7H-pyrrolo [2,3-d] pyrimidin-4-yl) -amino] -piperidin-1-yl} -3-oxo propionitrile, in free form or in a pharmaceutically acceptable salt form, for example, monitritrate (also called CP-690,550), or a compound as described in W02004052359 or W02005066156;

- mediators, eg activity agonists or modulators

S1P receptor, e.g. optionally phosphorylated FTY720 or an analog thereof, for example optionally phosphorylated 2-amino-2- [4- (3-benzyloxyphenylthio) -2-chlorophenyl] ethyl-1,3-propanediol 1- {4- [1- (4-cyclohexyl-3-trifluoromethyl-benzyloxy-imino) -ethyl] -2-ethyl-benzyl} -azetidine-3-one

carboxylic acid or pharmaceutically acceptable salts thereof;

immunosuppressive monoclonal antibodies, for example, leukocyte receptor monoclonal antibodies, for example, Blys / BAFF receptor, MHC, CD2, CD3, CD4, CD7, CD8, CD25, CD28, CD40, CD45, CD52, CD58, CD80, CD86 , IL-12 receptor, IL-17 receptor, IL-23 receptor or

its binders;

other immunomodulatory compounds, for example a recombinant binding molecule having at least a portion of the CTLA4 extracellular domain or a mutant thereof, for example at least a CTLA4 extracellular portion or a mutant thereof attached to a

non-CTLA4 protein sequence, e.g. CTLA4Ig (e.g., designated ATCC 68629) or a mutant thereof, e.g. LEA29Y;

- mediators, for example inhibitors of adhesion molecule activities, for example LFA-1 antagonists, ICAM-1 or 3 antagonists, VCAM-4 antagonists or VLA-4 antagonists,

- mediators, for example inhibitors, of FIM activity,

5-aminosalicylate (5-ASA) agents such as sulfasalazine, Azulfidine®, Asacol®, Dipentum®, Pentasa®, Rowasa®, Canasa®, Colazal®, for example mesalamine-containing drugs; for example mesalazine in combination with heparin;

- mediators, for example inhibitors, of TNF-alpha activity, for example, including antibodies that bind TNF-alpha, for example, influenza

ximab (Remicade®), thalidomide, lenalidomide,

- non-steroidal nitric oxide releasing anti-inflammatory drugs (NSAIDs), for example including COX inhibiting No donor drugs (CINOD);

- fosfordiesterase, for example mediators such as inhibitors of PDE4B activity,

- mediators, for example inhibitors, of caspase activity,

- mediators, eg agonists, of the protein G-coupled receptor GPBAR1,

- mediators, for example inhibitors, of ceramide activity

kinase,

"multifunctional anti-inflammatory" drugs (MFAIDs), for example cytosolic phospholipase A2 (cPLA2) inhibitors, such as glycosaminoglycan membrane-bound phospholipase A2 inhibitors;

antibiotics such as penicillins, cephalosporins, erythromycin, tetracyclines, sulfonamides such as sulfadiazine, sulfisoxazole; sulfo

nas, such as dapsone; pleuromutilins, fluoroquinolones, for example metronidazole, quinolones such as ciprofloxacin; levofloxacin; probiotic and commensal bacteria, for example, Lactobacillus, Lactobacillus reuteri;

antiviral drugs such as ribivirin, vidarabine, acyclovir, ganciclovir, zanamivir, oseltamivir phosphate, fanciclovir, atazanavir, amanta

dina, didanosine, efavirenz, foscarnet, indinavir, lamivudine, nelfinavir, ritonavir, saquinavir, stavudine, valaciclovir, valganciclovir, zidovudine.

Antiinflammatory drugs which are prone to be useful in combination with a compound of the present invention include, for example, non-steroidal antiinflammatory agents (NSAIDs) such as propionic acid derivatives (aminoprofen, benoxaprofen, bucloxic acid, carprofen, fenbufen , fenoprofen, fluprofen, flurbiprofen, ibuprofen, indoprofen, ketoprofen, miroprofen, naproxen, oxaprozine, pyrprofen, pranoprofen, suprofen, thiaprofenic acid, and thioxaprofen), acetic acid derivatives (indomethacin, acemetacin, clofenacenacoate, clofenacen, clofen phenclozic, fentiazac, furofenac, ibufenac, isoxepac, oxpi5 naco, sulindac, thiopinaco, tolmetin, zidomethacin, and zomepiraco), fenamic acid derivatives (fluphenamic acid, meclofenamic acid, mefenamic acid, niflumic acid and thiophenamic acid), thiophenamic acid (diflunisal and flufenisal), oxicans (isoxicam, piroxicam, sudoxicam and tenoxican) salicylates (acetyl salicylic acid, sulfasalazine) and pyrazolones (a10 pazone, bezpiperilone, feprazone, mofebutazone, oxyphenbutazone, phenylbutazone); cyclooxygenase-2 (COX-2) inhibitors such as celecoxib; phosphodiesterase type IV inhibitors (PDE-IV); chemokine receptor antagonists, especially CCR1, CCR2, and CCR3; cholesterol lowering agents such as HMG-CoA reductase inhibitors (lovastatin, simvas15 tatine and pravastatin, fluvastatin, atorvastatin, and other statins), sequestrants (cholestyramine and colestipol), nicotinic acid, fenofibric acid derivatives (genfibrozil, clofibrate, fenofibrate and benzafibrate), and probucol; anticholinergic agents such as muscarinic antagonists (ipratropium bromide); other compounds such as theophylline, sulfasalazine and aminosalicylates, for example 5-aminosalicylic acid and antirheumatic prodrugs thereof.

Antiallergic drugs that are prone to be useful in combination with a compound of the present invention include, for example, antihistamines (H1-histamine antagonists), for example bromopheniramine, chlorpheniramine, dexchlorpheniramine, triprolidine, clemastine, diphenhydramine, triphenylamine, triphenylamine, hydroxyzine, metdilazine, promethazine, trimeprazine, azatadine, cyproheptadine, antazoline, phenyramine pyrilamine, astemizole, terfenadine, loratadine, cetirizine, fexofenadine, decarbetoxyloratadine, such as non-steroidal meta-teroterol, non-steroidal anti-steroids isoetharine, albuterol, bitolterol, salmeterol and pyrbuterol), theophylline, sodium cromolin, atropine, ipratropium bromide, eukotriene antagonists (zafirlukast, montelukast, iralukast, pobilukast, SKB-106,203riene synthesis) BAY1005); bronchodilators, anti-asthmatics (mast cell stabilizers).

Anticancer drugs that are likely to be useful as a combination pair with a compound of the present invention, for example, include:

i. a steroid; for example prednisone.

ii. an adenosine kinase inhibitor; which targets, decreases or inhibits nucleobase, nucleoside, nucleotide and nucleic acid metabolism, such as 5-lodotubercidine, which is also known as 7H

pyrrolo [2,3-d] pyrimidin-4-amine, 5-iodo-7-p-D-ribofuranosyl.

iii. an adjuvant; which enhances the binding of 5-FU-TS as well as a compound that targets, decreases or inhibits alkaline phosphatase, such as leucovorin, Levaisol.

iv. adrenal cortex antagonist; which directs, decreases or inhibits adrenal cortex activity and alters the peripheral metabolism of corticosteroids

steroids, resulting in a decrease in 17-hydroxycorticosteroids such as mitotane.

v. an AKT pathway inhibitor; such as a compound that targets, decreases or inhibits Akt, also known as protein kinase B (PKB),

such as degueline, which is also known as 3H-bis [1'benzopyran [3,4-b: 6 ', 5'-e] pyran-7 (7aH) -one, 13,13a-dihydro-9,10 dimethyloxy-3,3-dimethyl- (7aS, 13aS); and triciribine, which is also known as 1,4,5,6,8-pentaazaacenaftilen-3-amine, 1,5-dihydro-5-methyl-1-β-D-ribofuranosyl.

saw. an alkylating agent; which causes DNA alkylation and results in fractures in DNA molecules as well as strand crosslinking

twins, thereby interfering with DNA replication and RNA transcription, such as nitrogen mustards, for example chlorambucil, chlormetine, cyclophosphamide, ifosfamide, melfalan, estramustine (Emcyt®); nitrosureas such as carmustine, fotemustine, lomustine, streptozocin (streptozotocin, STZ, Zanosar®), BCNU; Gliadel; dacarbazine, mechlorethamine, for example, as a hydrochloride, procarbazine, for example, as a hydrochloride, thiotepa, temozolomide (TEMODAR®), mitomycin, altretamine, busulfan, estramustine, uramustine. Cyclophosphamide may be administered, for example, in the form as it is marketed, for example under the trademark CYCLOSTIN®; and ifosfamide as HOLOXAN®.

vii. an angiogenesis inhibitor; which directs, decreases or inhibits the production of new blood vessels, for example, which directs methionine

aminopeptidase-2 (MetAP-2), inflammatory macrophage protein-1 (MIP)

1 alpha), CCL5, TGF-beta, lipoxygenase, cyclooxygenase, and topoisomerase, or which indirectly directs synthesis of p21, p53, CDK2 and collagen, for example, including fumagillin, which is known as mono ester [(3R, 4S, 105S, 6R) -5-methoxy-4 - [(2R, 3R) -2-methyl-3- (3-methyl-2-butenyl) oxiranyl] -1-oxaspiro [2,5] oct-6 2,4,6,8-decatetraenedioic acid-yl] (2E, 4E, 6E, 8E) - (9CI); chiconine, which is also known as 1,4-naphthalenedione, 5,8-dihydroxy-2 - [(1R) -1-hydroxy-4-methyl-3-pentenyl] - (9CI); tranylast, which is also known as benzoic acid, 2 - [[3- (3,4-dimethoxyphenyl) -1-oxo-2-propenyl] amino]; ursolic acid; suramin; bengamide or a derivative thereof, thalidomide, TNP470.

viii. an antiandrogen; which blocks the action of adrenal and testicular androgens that stimulate the growth of malignant normal and prostate tissue, such as nilutamide; bicalutamide (CASODEX®) which can

may be formulated for example as described in US4636505.

ix. an antiestrogen; which antagonizes the effect of estrogen at the estrogen receptor level, for example by including an aromatase inhibitor that inhibits estrogen production, ie the conversion of androstenedione and testosterone substrates to estrone and estradiol, respectively.

te, for example, including atamestane, exemestane, formestane, aminoglutethimide, rogletimide, pyridoglutethimide, trilostane, testolactone, ketoconazole, vorozole, fadrozole, anastrozole, letrozole, toremifene; bicalutamide; flutamide; tamoxifen, tamoxifen citrate; tamoxifen; fulvestrant; raloxifene, raloxifene hydrochloride. Tamoxifen may be, for example, administered in the form as it is marketed, for example NOLVADEX®; and raloxifene hydrochloride is marketed as EVISTA®. Fulvestrant may be formulated as described in US4659516 and is marketed as FASLODEX®. x. an antihypercalcemia agent; which is used to treat hypercalcemia, such as gallium nitrate hydrate (III); and disodium pamidronate.

xi an antimetabolite; which inhibits or disrupts DNA synthesis that results in cell death, such as folic acids, for example methotrexate,

pemetrexed, raltitrexed; purines, for example 6-mercaptopurine, cladribine, clofarabine; fludarabine, thioguanine (thioguanine), 6-thioguanine, pentostatin (deoxycoformycin); cytarabine; flexuridine; fluorouracil; 5-fluorouracil (5-FU), floxuridine (5-FUdR), capecitabine; gemcitabine; gemitabi hydrochloride; hydroxyurea (e.g., Hidrea®); DNA demethylating agents, such as 5-azacytidine and decitabine; edatrexate. Capecitabine and gemcitabine may be administered, for example, in commercial form, such as XELODA® and GEMZAR®.

xii. an apoptosis inducer; which induces the normal series of events in a cell leading to its death, for example by selectively inducing the X-linked mammalian inhibitor of apoptosis protein XIAP, or, for example, by down-regulating BCL-xL; such as ethanol, 2 - [[3- (2,3-dichlorophenoxy) propyl] amino]; gambogenic acid; embelin, which is also known as 2,5-cyclohexadiene-1,4-dione, 2,5-dihydroxy-3-undecyl- (9Cl); arsenic trioxide

k ■ 20 co.

xiii. an aurora kinase inhibitor; which directs, shortens, or inhibits later stages of the G2 / M checkpoint cell cycle all the way through the mitotic checkpoint and late mitosis; such as binuclein 2, which is also known as metanimidamide, N '- [1- (3-chloro

25 4-fluorophenyl) -4-cyano-1H-pyrazol-5-yl] -N, N-dimethyl- (9Cl).

xiv. A Bruton tyrosine kinase inhibitor (BTK); which directs, decreases or inhibits the development of human and murine B cells; such as terreic acid.

xv. a calcineurin inhibitor; which directs, decreases or inhibits the T-cell activation pathway such as cypermethrin, which is also known as cyclopropanecarboxylic acid, 3- (2,2-dichloroethenyl) -2,2-dimethyl-, cyano (3- phenoxyphenyl) methyl; deltamethrin, which is also known as cyclopropanecarboxylic acid, 3- (2,2-dibromoethenyl) -2,2-dimethyl- (S) -cyano (3-phenoxyphenyl) methyl ester, (1R, 3R); fenvalerate, which is also known as benzenoacetic acid, 4-chloro-α- (1-methylethyl) cyano (3-phenoxyphenyl) methyl ester; and Tirfostine 8; but excluding cyclosporine or FK506.

xvi. a CaM kinase II inhibitor; that directs, decreases or inhibits

CaM kinases; constituting a family of structurally related enzymes including phosphorylase kinase, light chain myosin kinase, and CaM kinases I to IV; such as 5-isoquinolinesulfonic acid, 4 - [(2S) -2 - [(5-isoquinolinylsulfonyl) methylamino] -3-oxo-3- (4-phenyl-1-piperazinyl) propyl] phenyl ester

(9CI); Benzenesulfonamide, N- [2 - [[[3- (4-chlorophenyl) -2-propenyl] methyl] amino] methyl] phenyl] -N- (2-hydroxyethyl) -4-methoxy.

xvii. a CD45 tyrosine phosphatase inhibitor; which directs, decreases or inhibits dephosphorylation pTyr regulated residues in Src family protein tyrosine kinases that assist in the treatment of a variety of

of inflammatory and immune disorders; such as phosphonic acid, [[2- (4-bromophenoxy) -5-nitrophenyl] hydroxymethyl].

xviii. a CDC25 phosphatase inhibitor; which directs, decreases or inhibits dephosphorylated over-expressed cyclin-dependent kinases in tumors; such as 1,4-naphthalenedione, 2,3-bis [(2-hydroethyl) thio].

xix a CHK kinase inhibitor; that directs, decreases or inhibits

overexpression of Bcl-2 antiapoptotic protein; such as debromoimenialdisine. Targets of a CHK kinase inhibitor are CHK1 and / or CHK2.

xx a controlling agent for regulating genistein, olomucine and / or tirphostins; such as daidzein, which is also known as 4H-1-

benzopyran-4-one, 7-hydroxy-3- (4-hydroxyphenyl); Iso-Olomoucine, and Tirfostina 1,

xxi. a cyclooxygenase inhibitor; for example including Cox-2 inhibitors; which directs, decreases or inhibits the enzyme Cox-2 (cyclooxygenase-2); such as 1H-indol-3-acetamide, 1- (4-chlorobenzoyl) -5-methoxy-2-one

methyl-N- (2-phenylethyl); Substituted 5-alkyl 2-arylaminophenylacetic acid and derivatives, for example celecoxib (CELEBREX®), rofecoxib (VIOXX®), etoricoxib, valdecoxib; or a 5-alkyl-2-arylaminophenylacetic acid, for example 5-methyl-2- (2'-chloro-6'-fluoroanilino) phenyl acetic acid, lumiracoxib; and celecoxib.

xxii. a cRAF kinase inhibitor; which directs, decreases or inhibits E-selectin over-regulation and molecule-1 induced vascular adhesion

5 TNF; such as 3- (3,5-dibromo-4-hydroxybenzylidene) -5-iodo-1,3-dihydroindole

2-one; and benzamide, 3- (dimethylamino) -N- [3 - [(4-hydroxybenzoyl) amino] -4-methylphenyl]. Raf kinases play an important role as extracellular signal regulatory kinases in cell differentiation, proliferation, and apoptosis. A target of a cRAF kinase inhibitor includes, but is not limited to, RAF1.

xxiii. a cyclin-dependent kinase inhibitor; which directs, decreases or inhibits cyclin-dependent kinase that plays a role in regulating the mammalian cell cycle; such as N 9 -isopropyl olomoucine; olomoucine; purvalanol B, which is also known as benzoic acid, 2-

chloro-4 - [[2 - [[(1 R) -1- (hydroxymethyl) -2-methylpropyl] amino] -9- (1-methylethyl) -9H-purin6-yl] amino] - (9CI); roascovitine; indirububin, which is also known as 2H-indol-2-one, 3- (1,3-dihydro-3-oxo-2H-indol-2-ylidene) -1,3-dihydro- (9CI) ; kenpaullone, which is also known as indole [3,2-d] [1] benzazepin6 (5H) -one, 9-bromo-7,12-dihydro- (9CI); purvalanol A, which is also known as 1-butanol, 2 - [[6 - [(3-chlorophenyl) amino] -9- (1-methylethyl) -9H-purin-2-yl] amino] -3-methyl -, (2 R) - (9 Cl); indirrubin-3'-mono-oxime. Cell cycle progression is regulated by a series of sequential events that include the activation and subsequent inactivation of cyclin-dependent kinases (Cdks) and cyclins. Cdks are a group of serine / threonine kinases that form active heterodimeric complexes by binding to their regulatory subunits, cyclins. Examples of targets of a cyclin-dependent kinase inhibitor include, but are not limited to, CDK, AHR, CDK1, CDK2, CDK5, CDK4 / 6, GSK3beta, and ERK.

xxiv. a cysteine protease inhibitor; which targets, decreases or inhibits cysteine protease which plays a vital role in mammalian cell turn and apotosis; such as 4-morpholinecarboxamide, N - [(1S) -3-fluoro-2-oxo-1- (2-phenylethyl) propyl] amino] -2-oxo-1- (phenylmethyl) ethyl]. xxv. a DNA interleaver; which binds to DNA and inhibits DNA, RNA, and protein synthesis; such as plicamycin, dactinomycin.

xxvi. a DNA strand breaker; which splits the DNA strand and results in inhibition of DNA synthesis, inhibition of RNA synthesis and

5 protein; such as bleomycin.

xxvii. an E3 ligase inhibitor; which directs, decreases or inhibits E3 ligase which inhibits the transfer of ubiquitin chains to proteins, marking them for proteasome degradation; such as N - ((3,3,3-trifluor-2-trifluoromethyl) propionyl) sulfanylamide.

xxviii. an endocrine hormone; that acting mainly on

Pituitary gland causes hormone suppression in males, the gross effect being a reduction of testosterone to castration levels; in females, ovarian estrogen and androgen synthesis being inhibited; such as leuprolide; megestrol, megestrol acetate.

xxix. compounds directing, decreasing or inhibiting the activity

receptor tyrosine kinases epidermal growth factor family (EGFR, ErbB2, ErbB3, ErbB4 as homoor heterodimers), such as compounds, proteins or antibodies that inhibit members of the EGF receptor tyrosine kinase family, e.g. EGF, ErbBI, ErbB2, ErbB3 and ErbB4 or EGF-binding or EGF-related Ligands, and are in particular those compounds, proteins or monoclonal antibodies specifically described in WO 9702266, for example, the compound of example 39, EP0564409, W09903854, EP0520722, EP0566226, EP0787722, EP0837063, US5747498, W09810767, W09730034, W09749688, W09738983, and especially W09630347, for example, a compound known as CP 358774, W09633980, known as ZD 1839; and WO 9503283, for example, a compound known as ZM105180, for example, including dual-action tyrosine kinase inhibitor (ErbB1 and ErbB2) Iapatinib (GSK572016), for example, lapatinib ditosylate; panituzumab, trastuzumab (HERCEPTIN®), cetuximab, iressa, OSI-774, CI-1033, EKB-569, GW-2016, E1.1, E2.4, E2.5, E6.2, E6.4, E2. 11, E6.3 or E7.6.3, 7H-pyrrolo [2,3-d] pyrimidine derivatives which are, for example, described in WO003013541, erlotinib, gefitinib. Erlotinib may be administered as marketed, for example, TARCEVA®, and gefitinib as IRESSA®, human monoclonal antibodies against epidermal growth factor receptor including ABX-EGFR.

xxx an EGFR inhibitor, PDGFR tyrosine kinase; as ini

EGFR kinase inhibitors including tirfostine 23, tirfostine 25, tirfostine 47, tirfostine 51 and tirfostine AG 825; 2-propenamide, 2-cyano-3- (3,4-dihydroxyphenyl) -N-phenyl- (2E); tirfostine Ag 1478; Yavendustine A; 3-pyridinoacetonitrile, α [(3,5-dichlorophenyl) methylene] -> (aZ); An example of an EGFR, PDGFR tyrosine kinase inhibitor, for example, includes typhostin 46, PDGFR tyrosine kinase inhibitor including typhostin 46. Targets of an EGFR kinase inhibitor include guanylyl cyclase (GC-C) HER2, EGFR, PTK and tubulin.

xxxi. a farnesyltransferase inhibitor; which directs, decreases or inhibits Ras protein; such as α-hydroxypharnesylphosphonic acid; buta acid

Noic, 2 - [[(2S) -2 - [[(2S, 3S) -2 - [[(2R) -2-amino-3-mercaptopropyl] amino] -3-methylpentyl] oxy] -1-ester oxo-3-phenylpropyl] amino] -4- (methylsulfonyl) -1,1-methylethyl (2S); manumycin A; L-744,832 or DK8G557, tipifarnib (R115777), SCH66336 (canvasfamib), BMS-214662.

xxxii. a Flk-1 kinase inhibitor; which directs, decreases or inhibits Flk-1 tyrosine kinase activity; such as 2-propenamide, 2-cyano-3- [4-

hydroxy-3,5-bis (1-methylethyl) phenyl] -N- (3-phenylpropyl) - (2E). A target of a Flk-1 kinase inhibitor includes, but is not limited to, KDR.

xxxiii. a glycogen kinase-3 synthase inhibitor (GSK3); which directs, decreases or inhibits glycogen kinase-3 synthase (GSK3); as indi

rubin-3'-mono-oxime. Glycogen Kinase-3 Syntase (GSK-3; protein tau kinase I), a ubiquitously expressed highly conserved serine / threonine kinase protein, is involved in the signal transduction cascades of multiple cell processes, which is a protein kinase that has been shown to involved in regulating a diverse array of cellular functions, including protein synthesis, cell proliferation, cell differentiation, microtubule assembly / disassembly, and apoptosis.

xxxiv. a histone deacetylase inhibitor (HDAC); which inhibits histone deacetylase and has antiproliferative activity; such as compounds described in WO00222577, especially N-hydroxy-3- [4 - [[(2-hydroxyethyl) [2- (1H-indol-3-yl) ethyl] -amino] methyl] phenyl] -2E-2 -propenamide, and N-hydroxy-3- [4 - [[[[2- (2-methyl-1H-indol-3-yl) ethyl] amino] methyl] phenyl] -2E-2-propenamide and salts Pharmaceutically acceptable thereof; suberoylanilide hydroxamic acid (SAHA); [4- (2-Amino-phenylcarbamoyl) -benzyl] -carbamic acid pyridine-3-ylmethyl ester and derivatives thereof; butyric acid, pyroxamide, trichostatin A, oxanflatin, apicidine, depsipeptide; depudecin; trapoxin, HCl toxin which is also known as cyclo [L-alanyl-D-alanyl - ((S, 2S) - (10 amino - (- oxo-oxiraneoctanoyl-D-prolyl] (9CI); sodium phenylbutyrate, bishidroxamic acid Trichostatin A, BMS-27275, pyroxamide, FR901228, valproic acid.

xxxv. An HSP90 inhibitor; which directs, decreases or inhibits the intrinsic ATPase activity of HSP90; degrades, directs, decreases or inhibits client HSP90 proteins via the ubiquitin proteasome pathway. Compounds that target, decrease or inhibit HSP90 intrinsic ATPase activity are especially compounds, proteins or antibodies that inhibit HSP90 ATPase activity, for example, 17-allylamino, 17-demethoxygeldanamycin (17AAG), a geldanamycin derivative; other geldanamycin related compounds; radicicol and HDAC inhibitors. Other examples of an HSP90 inhibitor include geldanamycin, 17-demethoxy-17- (2-propenylamino). Potential indirect targets of an HSP90 inhibitor include FLT3, BCR-ABL, CHK1, CYP3A5 * 3 and / or NQ01 * 2. Nilotinib is an example of a BCR-ABL tyrosine kinase inhibitor.

xxxvi. a 1-kappa B-alpha kinase (IKK) inhibitor; that directs,

decreases or inhibits NF-κB, such as 2-propenenitrile, 3 - [(4-methylphenyl) sulfonyl] - (2E).

xxxvii. an insulin receptor tyrosine kinase inhibitor; modulating the activities of phosphatidylinositol 3-kinase, a protein associated with

microtubule, and S6 kinases; such as hydroxyl-2-naphthalenylmethylphosphonic acid, LY294002.

xxxviii. a c-Jun N-terminal kinase (JNK) kinase inhibitor; which directs, decreases or inhibits Jun N-terminal kinase; such as pyrazoleanchair and / or epigallocatechin gallate. Jun N-terminal kinase (JNK), a serine-directed protein kinase, is involved in the phosphorylation and activation of c-Jun and ATF2 and plays a significant role in metabolism, growth, cell differentiation, and apoptosis. A target for a JNK kinase inhibitor includes, but is not limited to, DNMT.

xxxix. a microtubule binding agent; which acts by disrupting the microtubular network that is essential for mitotic cell function and interphase; such as vinca alkaloids, for example vinblastine, vinblastine sulfate;

Vincristine, vincristine sulfate; vindesine; vinorelbine; taxanes, such as taxanes, for example docetaxel; paclitaxel; discodermolids; cochicin, epothilones and derivatives thereof, for example epothilone B or a derivative thereof. Paclitaxel is marketed as TAXOL®; docetaxel as TAXOTERE®; vinblastine sulfate as VINBLASTIN R.P®; and vincristine sulfate as FARMISTIN®. Also included are generic paclitaxel forms as well as various paclitaxel dosage forms. Generic forms of paclitaxel include, but are not limited to, betaxolol hydrochloride. Several paclitaxel dosage forms include, but are not limited to, albumin nanoparticle paclitaxel marketed as ABRA'20®; ONXOL®, CYTOTAX®. Discodermolide may be obtained, for example, as described in US5010099. Also included are Epotholine derivatives which are described in US 6194181, WO98 / 0121, WO9825929, WO09808849, WO9943653, WO9822461 and WO00031247. Especially preferred is Epotholine A and / or B.

xl. a mitogen activated protein (MAP) kinase inhibitor;

which targets, decreases or inhibits mitogen-activated protein such as benzenesulfonamide, N- [2 - [[[[3- (4-chlorophenyl) -2-propenyl] methyl] amino] methyl] phenyl] N- (2-hydroxyethyl) -4-methoxy. Mitogen-activated protein kinases (MAP) are a group of serine / threonine protein kinases that are activated in response to a variety of extracellular stimuli and mediate signal transduction from the cell surface to the nucleus. They regulate various physiological and pathological cellular phenomena, including inflammation, apoptotic cell death, oncogenic transformation, tumor cell invasion, and metastasis.

xli. an MDM2 inhibitor; which directs, decreases or inhibits the interaction of MDM2 and the p53 tumor suppressor; such as trans-4-iodine, 4'boranyl-chalcone.

5 xi. a MEK inhibitor; that directs, decreases or inhibits activity

MAP MEK kinase kinase; such as sorafenib, for example Nexavar® (sorafenib tosylate), butanedinitrile, bis [amino [2-aminophenyl) thio] methylene]. A target of a MEK inhibitor includes, but is not limited to, ERK. An indirect target of a MEK inhibitor includes, but is not limited to, cyclin D1. Hi. a matrix metalloproteinase inhibitor inhibitor

(MMP); which targets, decreases or inhibits a protease enzyme class that selectively catalyzes the hydrolysis of polypeptide bonds including the enzymes MMP-2 and MMP-9 that are involved in promoting loss of tissue structure around tumors and facilitating growth. of tumor, angiogenesis, and metastasis such as actinonine, which is also known as butanediamine, N-4-hydroxy-N1 - [(1S) -1 - [[(2S) -2- (hydroxymethyl)

1-pyrrolidinyl] carbonyl] -2-methylpropyl] -2-pentyl-, (2R) - (9Cl); epigallocatechin gallate; peptidomimetic and non-peptidomimetic collagen inhibitors; tetracycline derivatives, for example hydroxide mato batimastat peptidomimetic inhibitor; and their orally bioavailable analogues marimastat, prinomastat, metastat, neovastat, tanomastat, TAA211, BMS-279251, LADRAM 12-9566, MMI270B or AAJ996. A target of an MMP inhibitor includes, but is not limited to, polypeptide deformylase.

xliv. an NGFR tyrosine kinase inhibitor; which directs, decreases or inhibits nerve development factor-dependent p140c‘trk tyrosine phosphorylation; such as tirfostine AG 879. Targets of an NGFR tyrosine kinase inhibitor include, but are not limited to, HER2, FLK1, FAK, TrkA, and / or TrkC. An indirect target inhibits RAF1 expression.

xlv. a p38 MAP kinase inhibitor, including a SAPK2 / p38 kinase inhibitor; which targets, decreases or inhibits p38-MAPK which is a member of the MAPK family such as phenol, 4- [4- (4-fluorophenyl) -5- (4-pyridinyl) 1H-imidazol-2-yl]. An example of an SAPK2 / p38 kinase inhibitor includes, but is not limited to, benzamide, 3- (dimethylamino) -N- [3 - [(4-hydroxybenzoyl) amino] -4-methylphenyl]. One member of the MAPK family is a tyrosine phosphorylation-activated serine / threonine kinase and threonine residues. This kinase is phosphorylated and activated by many cell strains and inflammatory stimuli thought to be involved in the regulation of important cellular responses such as apoptosis and inflammatory reactions.

xlvi. a p56 tyrosine kinase inhibitor; which directs, decreases or inhibits p56 tyrosine kinase which is an enzyme that is a lymphoid-specific src family tyrosine kinase critical for T cell development and activation; such as damacantal, which is also known as 2-anthracenecarboxaldehyde, 9,10-dihydro-3-hydroxy-1-methoxy-9,10-dioxo, Tirfostine 46. A target of a p56 tyrosine kinase inhibitor includes, but It's not limited to, Lck. Lck is associated with the CD4, CD8 cytoplasmic domains and the IL-2 receptor beta chain, and is believed to be involved in the most premature stages of TCR-mediated T cell activation.

xlvii. a PDGFR tyrosine kinase inhibitor; directing, decreasing or inhibiting the activity of C-kit receptor tyrosine kinases (part of the PDGFR family), such as directing, decreasing or inhibiting the activity of the c-kit receptor tyrosine kinase family, especially in

- 20 c-kit receiver. Examples of targets of a PDGFR tyrosine kinase inhibitor include, but are not limited to, PDGFR, FLT3 and / or c-kit; such as tirfostine AG 1296; tirfostine 9; 1,3-butadiene-1,1,3-tricarbonitrile, 2-amino-4- (1H-indol-5-yl); N-phenyl-2-pyrimidine amine derivative, e.g., imatinib, IRESSA®. PDGF plays a central role in regulating cell proliferation, chemotaxis, and survival in normal cells as well as in various disease states such as cancer, atherosclerosis, and fibrotic disease. The PDGF family is composed of dimeric isoforms (PDGF-AA, PDGF-BB, PDGF-AB, PDGF-CC, and PDGF-DD) by exerting their cellular effects differentially by binding to the two receptor tyrosine kinases. 30 PDGFR-α and PDGFR-β have molecular masses of 170 and 180 kDa, respectively.

xviii. a phosphatidylinositol 3 kinase inhibitor; which directs, decreases or inhibits P1 kinase; such as wortmannin, which is also known as 3H-Hole [4,3,2-de] indene [4,5-h] -2-benzopyran-3,6,9-trione, 11- (acetyloxy) -1, 6b, 7,8,9a, 10,11,11.b-octahydro-1- (methoxymethyl) -9a, 11b-dimethyl-, (1S, 6bR, 9aS, 11 R, 11 bR) - (9CI ); 8-phenyl-2- (morpholin-4-yl) -chromen-4-one; quercetin, quercetin dihydrate. Pl 3-kinase activity has been shown to increase in response to various hormonal and growth factor stimuli, including insulin, platelet-derived growth factor, insulin-like growth factor, epidermal growth factor, colony stimulating factor, and hepatocyte growth, and has been implicated in processes related to cell growth and transformation. An example of a target of a phosphatidylinositol 3-kinase inhibitor includes, but is not limited to, Pi3K.

xlix. a phosphatase inhibitor; which directs, decreases or inhibits phosphatase; such as chanting acid; cantharidin; and L-leucinamide, N- [4- (2-15 carboxyethenyl) benzoyl] glycyl-L-α-glutamyl- (E). Phosphatases remove the phosphorylated group and restore the protein to its original dephosphorylated state. Consequently, the phosphorylation and dephosphorylation cycle can be considered an "on-off" molecular switch.

I. platinum agent; which contains platinum and inhibits DNA synthesis by forming interphase and intra-strand cross-linking of DNA molecules; such as carboplatin; cisplatin; oxaliplatin; cisplatin; satraplatin and platinum agents such as ZD0473, BBR3464. Carboplatin may be administered, for example, in the form as it is marketed, for example, CARBOPLAT®; and oxaliplatin as ELOXATIN®.

read it a protein phosphatase inhibitor, including a PP1 inhibitor

and PP2 and a tyrosine phosphatase inhibitor; which directs, decreases or inhibits protein phosphatase. Examples of a PP1 and PP2A inhibitor include cantharidic acid and / or cantharidine. Examples of a tyrosine phosphatase inhibitor include, but are not limited to, L-β-bromotetramisol oxalate; 2 (5H) furanone, 4-hydroxy-5- (hydroxymethyl) -3- (1-oxohexadecyl) -, (5R); and benzylphosphonic acid.

The term "a PP1 or PP2 inhibitor" as used herein refers to a compound that targets, decreases or inhibits Ser / Thr protein phosphatases. Type I phosphatases, which include PP1, may be inhibited by two thermostable proteins known as inhibitor-1 (1-1) and inhibitor-2 (I-2). They preferably dephosphorylate a phosphorylase kinase subunit. Fos5 type II phosphates are spontaneously subdivided into active phosphatase (PP2A), CA2 + -dependent (PP2B), and Mg2 + -dependent (PP2C) classes.

The term "tyrosine phosphatase inhibitor" as used herein refers to a compound that targets, decreases or inhibits tyrosine phosphatase. Protein tyrosine phosphatases (PTPs) are relatively recent additions to the phosphatase family. They remove the phosphate groups from the phosphorylated tyrosine residues of proteins. PTPs exhibit diverse structural characteristics and play important roles in regulating cell proliferation, cell differentiation, adhesion and motility, and cytoskeletal function. Examples of targets of a tyrosine phosphatase inhibitor include, but are not limited to, alkaline phosphatase (ALP), heparanase, PTPase, and / or prostatic acid phosphatase.

III. a PKC inhibitor and a PKC delta kinase inhibitor: The term "a PKC inhibitor" as used herein refers to a compound that directs, decreases or inhibits protein C kinase as well as its isozymes.

► 20 Protein kinase C (PKC), a ubiquitous phospholipid-dependent enzyme, is involved in signal transduction associated with cell proliferation, differentiation, and apoptosis. Examples of a target of a PKC inhibitor include, but are not limited to, MAPK and / or NF-capaB. Examples of a PKC inhibitor include, but are not limited to, 1-h-pyrrol-2,5-dione, 3- [1- [3- (dimethyl25 amino) propyl] -1H-indol-3-yl] -4- (1H-indol-3-yl); bisindolylmaleimide IX; sphingosine, which is known as 4-octadecene-1,3-diol 2-amino-, (2S, 3R, 4E) (9CI); staurosporine, which is known as 9,13-epoxy-1H, 9H-diindolo [1,2,3-gh: 3,2,2 ', 1'-1m] pyrrolo [3,4-j] [1,7] benzodiazonin-1-one, staurosporine derivatives such as described in EP0296110, for example midostauri30 na; 2,3,10,11,12,13-hexahydro-10-methoxy-9-methyl-11- (methylamino) - (9S, 10R, 11R, 13R) - (9CI); tirfostine 51; and hypericin, which is also known as phenanthro [1,10,9,8-opqra] perylene-7,14-dione, 1,3,4,6,8,13-hexahydroxy-10,11-dimethyl, stereoisomer (6CI, 7CI, 8CI, 9CI), UCN-01, safingol, BAY 43-9006, bryostatin 1, periphosine; llmophosine; RO 318220 and RO 320432; RO 6976; Isis 3521; LY333531 / LY379196. The term "a PKC delta kinase inhibitor" as used herein refers to a compound that targets, decreases or inhibits PKC delta isozymes. The delta isozyme is a conventional PKC isozyme and is Ca 2+ -dependent. An example of a PKC delta kinase inhibitor includes, but is not limited to, Rottlerin, which is also known as 2-Propen-1-one, 1- [6 - [(3-acetyl-2,4,6-tri -hydroxy-5-methylphenyl) methyl] -5,7-dihydroxy-2,2-dimethyl-2H-1-benzopyran-8-yl] -3-phenyl-, (2E) - (9Cl).

Hi. a polyamine synthesis inhibitor; that directs, decreases or

inhibits spermidine from polyamines; such as DMFO, which is also known as (-) - 2-difluoromethylornithine; N1, N12-diethylpermin 4HCl. The spermidine and spermine polyamines are of vital importance for cell proliferation, although their precise mechanism of action is unclear. Tumor cells 15 have an altered polyamine homeostasis reflected by increased activity of biosynthetic enzymes and elevated general polyamine backgrounds.

Iiv. a proteasome inhibitor; which targets, decreases or inhibits proteasome, such as aclacinomycin A; glyiotoxin; PS-341; MLN 341; bortezomib; Velcade. Examples of targets of a proteasome inhibitor include, but are not limited to, 0 (2) (-) - generating NADPH oxidase, NFapapaB, and / or farnesyltransferase, geranyltransferase I.

Iv. a PTP1B inhibitor; which targets, decreases or inhibits PTP1B, a protein tyrosine kinase inhibitor; such as L-leucinamide, N- [4- (2-carboxyethenyl) benzoyl] glycyl-L-α-glutamyl -, (E).

I saw. a protein tyrosine kinase inhibitor including an inhibitor

SRC family tyrosine kinase; a Syk tyrosine kinase inhibitor; and a JAK-2 and / or JAK-3 tyrosine kinase inhibitor;

The term "a protein tyrosine kinase inhibitor" as used herein refers to a compound that targets, decreases or inhibits protein tyrosine kinases. Protein tyrosine kinases (PTKs) play a key role in regulating cell proliferation, differentiation, metabolism, migration, and survival. They are classified as receiver PTKs and non-receiver PTKs. Receptor PTKs contain a single polypeptide chain with a transmembrane segment. The extracellular end of this segment contains a high affinity ligand binding domain, while the cytoplasmic end comprises the catalytic nucleus and regulatory sequences. Examples of targets of a tyrosine kinase inhibitor include, but are not limited to, ERK1, ERK2, Bruton Tyrosine Kinase (Btk), JAK2,% ERK, PDGFR, and / or FLT3. Examples of indirect targets include, but are not limited to, TNFalpha, NO, PGE2, IRAK, iNOS, ICAM-1, and / or E-selectin. Examples of a tyrosine kinase inhibitor included

10 in, but are not limited to, tirfostine AG 126; tirfostine Ag 1288; tirfostine Ag 1295; geldanamycin; and genistein.

Non-receptor tyrosine kinases include members of the Src, Tec, JAK, Fes, Abi, FAK, Csk, and Syk families. They are located in the cytoplasm as well as in the nucleus. They exhibit distinct kinase regulation, substrate phosphorylation, and function. Deregulation of these kinases has also been linked to various human diseases.

The term "an SRC family tyrosine kinase inhibitor" as used herein refers to a compound that targets, decreases or inhibits SRC. Examples of an SRC family tyrosine kinase inhibitor include, but are not limited to,

- 20 are not limited to, PP1, which is also known as 1 H-pyrazolo [3,4-

d] pyrimidin-4-amine, 1- (1,1-dimethylethyl) -3- (1-naphthalenyl) - (9 Cl); and PP2, which is also known as 1H-Pyrazolo [3,4-d] pyrimidin-4-amine, 3- (4-chlorophenyl) -1- (1,1-dimethylethyl) - (9CI).

The term "a Syk tyrosine kinase inhibitor" as used herein refers to a compound that targets, decreases or inhibits Syk. Examples of targets for a Syk tyrosine kinase inhibitor include, but are not limited to.

a, Syk, STAT3, and / or STAT5. An example of a Syk tyrosine kinase inhibitor includes, but is not limited to, piceatanol, which is also known as 1,2-benzenediol, 4 - [(1 E) -2- (3,5-dihydroxyphenyl) ethenyl ] - (9CI).

The term "a Janus inhibitor (JAK-2 and / or JAK-3) tyrosine kina

if, "as used herein, refers to a compound that targets, decreases or inhibits janus tyrosine kinase. Janus tyrosine kinase inhibitor is shown to have antileukemic agents with antithrombotic, antiallergic and immunosuppressive properties. Targets for a JAK-2 inhibitor and / or JAK-3 tyrosine kinase include, but are not limited to, JAK2, JAK3, STAT3 An indirect target of a JAK-2 and / or JAK-3 tyrosine kinase inhibitor includes, but is not limited to, 5 CDK2. A JAK-2 and / or JAK-3 tyrosine kinase inhibitor include, but are not limited to, Tirfostin AG 490 and 2-naphthyl vinyl ketone.

Compounds that direct, decrease or inhibit the activity of c-Abl family members and their gene fusion products, for example, include PD180970; AG957; or NSC 680410.

Ivii. a retinoid; that directs, decreases or inhibits reti receptors

noide-dependent; such as isotretinoin, tretinoin, allitretinoin, bexarotene.

Iviii. an RNA II polymerase elongation inhibitor; which directs, decreases or inhibits insulin-stimulated nuclear and cytosolic p70S6 kinase in CHO cells; directs, decreases or inhibits polymerase transcription

RNA II which may be casein kinase II dependent; and directs, decreases or inhibits germ vesicle disruption in bovine oocytes; such as 5,6-dichloro-1-beta-D-ribofuranosylbenzimidazole.

Ivix a serine / threonine kinase inhibitor; which inhibits serine / threo kinase kinases; such as 2-aminopurine. An example of a serine / threonine kinase inhibitor target includes, but is not limited to, dsRNA-dependent protein kinase (PKR). Examples of indirect targets of a serine / threonine kinase inhibitor include, but are not limited to, MCP-1, NF-kapB, elF2alpha, COX2, RANTES, IL8, CYP2A5, IGF-1, CYP2B1, CYP2B2, CYP2H1, 25 AI -1, HIF-1, erythropoietin, and / or CYP1A1.

Ix. a sterol biosynthesis inhibitor; which inhibits the biosynthesis of sterols such as cholesterol; such as terbinadine. Examples of targets for a sterol biosynthesis inhibitor include, but are not limited to, squalene epoxidase, and CYP2D6.

Ixi. a topoisomerase inhibitor; including a topoi inhibitor

somerase I and a topoisomerase II inhibitor. Examples of a topoisomerase I inhibitor include, but are not limited to, topotecan, gimatecan, irinotecan, camptotecan and their analogs, 9-nitrocamptothecin and macromolecular camptothecin conjugate PNU-166148 (compound A1 in WO09917804); 10-hydroxycamptothecin, for example the acetate salt; idarubicin, for example the hydrochloride; irinotecan, for example the hydrochloride; eto5 poside; teniposide; topotecan, topotecan hydrochloride; doxorubicin; epirubicin, epirubicin hydrochloride; mitoxantrone, mitoxantrone, for example hydrochloride; daunorubicin, daunorubicin hydrochloride, valrrubicin, dasatinib (BMS-354825). Irinotecan may be administered, for example, in the form as it is marketed, for example under the trademark CAMPTO10 SAR®. Topotecan may be administered, for example, in the form as it is marketed, for example under the trademark HYCAMTIN®. The term "topoisomerase II inhibitor" as used herein includes, but is not limited to, anthracyclines, such as doxorubicin, including liposomal formulation, e.g., CAELYX®, daunorubicin, including liposomal formulation, e.g., DAUNOSOME®. epirubicin, idarubicin and nemorubicin; the anthraquinones mitoxantrone and losoxantrone; and the etoposide and teniposide podophyllotoxins. Etoposide is marketed as ETOPOPHOS®; teniposide as VM 26-BRISTOL®; doxorubicin such as ADRIBLASTIN® or ADRIAMYCIN®; epirubicin as FARMORUBICIN®, idarubicin as ZAVE

- 20 DOS®; and mitoxantrone as NOVANTRON®.

Ixii. VEGFR tyrosine kinase inhibitor; which directs, decreases and / or inhibits known angiogenic growth factors and cytokines implicated in the modulation of normal and pathological angiogenesis. The VEGF family (VEGF-A, VEGF-B, VEGF-C, VEGF-D) and their corresponding receptor tyrosine kinases [VEGFR-1 (Flt-1), VEGFR-2 (Flk-1, KDR), and VEGFR-3 (Flt-4)] play a supreme and indispensable role in regulating the multiple facets of angiogenic and lymphangiogenic processes. An example of a VEGFR tyrosine kinase inhibitor includes 3- (4-dimethylaminobenzylidenyl) -2-indolinone. Compounds that direct, decrease or inhibit VEGFR activity are especially compounds, proteins or antibodies that inhibit VEGF receptor tyrosine kinase, inhibit a VEGF receptor or bind to VEGF, and are in particular those generic monoclonal compounds, proteins or antibodies. and specifically described in WO9835958, for example 1- (4-chloroanilino) -4- (4-pyridylmethyl) phthalazine or a pharmaceutically acceptable salt thereof, for example succinate, or in W00009495, W00027820, W00059509, W09811223, W00027819 and 5 EP0769947; for example those as described by M. Prewett et al in Cancer Research 59 (1999) 5209-5218, by F. Yuan et al in Proc. Natl. Acad. Know. USA, vol. 93, p. 14765-14770, ten. 1996, by Z. Zhu et al in Cancer Res. 58,1998,3209-3214, and by J. Mordenti et al in Toxicologic Pathology, Vol. 27, no. 1, pages 14-21.1999; in W00037502 and WO9410202; Angiostatin, described by M., S. CReiIIy et al., Cell 79,1994,315-328; Endostatin described by M. S. O'Reilly et al, Cell 88,1997,277-285; anthranilic acid amides; ZD4190; ZD6474 (vandetanib); SU5416; SU6668; or antiVEGF antibodies or antiVEGF receptor antibodies, for example RhuMab (bevacizumab). By antibody is meant intact monoclonal antibodies, polyclonal antibodies, multispecific antibodies formed of at least 2 intact antibodies, and antibody fragments so long as they exhibit the desired biological activity. An example of a VEGF-R2 inhibitor, for example, includes axitinib.

Iiiii. a gonadorelin agonist such as abarelix, goserelin, goserelin acetate,

IX. a compound that induces cell differentiation processes such as retinoic acid, alpha-, gamma- or 8-tocopherol or alpha-, gamma- or 8-tocotrienol.

Ixv. a bisphosphonate, for example, including etridonic, clodronic, tiludronic, pamidronic, alendronic, ibandronic, risedronic and zoledronic acid.

Ixvi. a heparanase inhibitor that prevents degradation of heparan sulfate, e.g. PI-88.

Ixvii. a biological response modifier, preferably alinfocin or interferones, for example interferon alfa.

VIII. a telomerase inhibitor, for example telomestatin.

Ixix. mediators such as catechol-O-methyltransferase inhibitors, for example entacapone.

Ixx: ispinesib, permetrexed (Alimta®), sunitinib (SU11248), diethylstiibestro (DES) 1 BMS224818 (LEA29Y).

Ixxi. somatostatin or a somatostatin analogue such as octreotide (Sandostatin® or Sandostatin LAR®).

Ixxii. growth hormone receptor antagonists such as pegvisomant, filgrastim or pegfilgrastim, or interferon alfa:

Ixxiii. monoclonal antibodies, for example useful for treating leukemia (AML), such as alemtuzumab (Campath®), rituximab / Ritu10 xan®), gentuzumab, (ozogamycin, Mylotarg®), and pratuzumab.

Ixxiv. altretamine, ansacrine, asparaginase (Elspar®), denileucin diftitox, masoprocol, pegaspargase.

Ixxv. a phosphodiesterase inhibitor, for example anagrelide (Agrylin®, Xagrid®).

15 Ixxvi. a cancer vaccine, such as MDX-1379.

Treatment of cancer with a compound of the present invention, optionally in combination with an anticancer drug as indicated herein, may be associated with radiotherapy. Cancer treatment with a compound of the present invention, optionally in combination with an anticancer drug, may be a second line treatment, for example, following treatment with another anticancer drug or other cancer therapy.

Anesthetics which are prone to be useful as a combination pair with a compound of the present invention, for example, include ethanol, bupivacaine, chloroprocaine, levobupivacaine, lidocaine, mepivacaine, procaine, ropivacaine, tetracaine, desflurane, isoflurane, ketamine, propofol, sevoflurane, codeine, fentanyl, hydromorphone, marcaine, meperidine, methadone, morphine, oxycodone, remifentanil, butfanol, nalbuphine, tramadol, benzocaine, dibucaine, ethyl chloride, xylocaine, and phenazopyridine.

30 Antidiarrheal drug substances that are prone to be

Useful as a combination pair with an IBD agent or agent of the present invention, for example, include diphenoxylate, loperamide, codeine. If a compound of the present invention is administered in combination with other drug substance dosages of the second co-administered drug substance will, of course, vary depending upon the type of co-employed drug, the specific drug employed, the condition being treated, as in the case of a drug. compound of the present invention. In general dosages similar to those as provided by the second drug provider may be appropriate.

The chemical names of the compounds of the present invention as indicated herein are copied from ISIS, version 2.5 (AutoNom Name 2000). Chemical names may be derived from second drug substances and other substances on the Internet, for example, through a research program such as SCI FINDER.

In the following Examples all temperatures given are in ° Celsius (abbreviated as °). The following abbreviations are used:

DMAP 4- (dimethylamino) pyridine DMF N, N-dimethylformamide EtOAc ethyl acetate rac-BINAP rac-2,2'-Bis (diphenylphosphino) -1,1'-binaphthyl Pd (dba) 2 Bis (dibenzylidenoacetone) palladium PPA anhydride 1-propanophosphonic acid cyclic acid at room temperature THF tetrahydrofuran t-Bu tert-butyl EXAMPLE 1 1- (4-tert-Butyl-benzenesulfonyl) -5-chloro-1 H -indole-2-carbo acid

xylene (Compound 1 from Table 1)

A) 1- (4-TERC-BUTIL-BENZENOSULFONIU ACID ETHYL ESTER

5-CHLOR-1H-INDOL-2-CARBOXYL

244 mg of 18-crown-6 are added to 12 ml of a solution of potassium tert-butoxide (1 M in THF) and to the mixture obtained a solution of

2 g of 5-chloro-1H-indole-2-carboxylic acid ethyl ester in 24 ml of THF is added dropwise. The obtained mixture is cooled to 0 ° C (15 minutes), treated with a solution of 2.08 g of 4-tert-butylbenzenesulfonyl chloride in 12 ml THF, stirred for 16 hours at RT and the obtained mixture is diluted with EtOAc. The obtained organic phase is washed with 0.1 N HCl brine, dried, and from the obtained mixture the solvent is evaporated. The evaporation residue obtained is subjected to chromatography.

1- (4-tert-Butyl-benzenesulfonyl) -5-chloro-1H-indol-2-carboxylic acid ethyl ester is obtained.

B) 1- (4-TERC-BUTIL-BENZENOSULPHONIU-5-CHLOR-1 H-INDOL ACID

2-CARBOXYLIC

To a solution of 1.05 g of 1- (4-tertiary acid ethyl ester)

butylbenzenesulfonyl) -5-chloro-1 H -indole-2-carboxylic acid in 40 ml THF, a solution of 318 mg LiOH x H2 O in 40 ml HO is added. The obtained mixture is stirred for 18 hours at RT and the obtained organic phase is evaporated. The obtained aqueous phase is washed with CHCl3, the pH of the obtained mixture is adjusted to pH = 4 by addition of 6 N HCl and the obtained mixture is extracted with EtOAc. The obtained combined organic layers are dried, the obtained solvent mixture is evaporated and the obtained evaporation residue is subjected to RP-HPLC. 1- (4-tert-Butyl-benzenesulfonyl) -5-chloro-1H-indol-2-carboxylic acid is obtained, mp 198-202.

Similarly as described in Example 1, but using the ma

Suitable starting materials, the compounds of Examples 2 to 10 as shown and characterized in Table 1 are obtained.

EXAMPLE 11

1- (4-tert-Butyl-benzenesulfonyl) -5-chloro-1 H -indole-2-carboxylic acid (compound of example 11 in TABLE 1)

A) 1- (4-TERC-BUTIL-BENZENOSSULFONILV1 H-INDOL

To a solution of 300 mg indole in 10 ml DMF1 151 mg NaH (60% in mineral oil) and 596 mg 4-tert-butyl is added benzene sulfonyl chloride. The obtained mixture is stirred for 18 hours in RT 30 poured into H 2 O and extracted with EtOAc. The obtained combined organic layers are dried, the obtained solvent mixture is evaporated and the evaporation residue is precipitated from EtOH. 1- (4-tert-Butyl-benzenesulfonyl) -1H-indole is obtained.

B) 1- (4-TERC-BUTIL-BENZENQSULFONYL-1H-INDOL-2-CARBOXYLIC ACID

460 μΙ n-BuLi (1.6 N hexane solution) is added over 10 minutes dropwise to a solution of 105 μΙ diisopropylamine in 2 ml dry THF at -78 °. The obtained solution is kept warming to 0 ° C (30 minutes) and is treated with a solution of 1- (4-tert-butylbenzenesulfonyl) -1 H -indole 200 mg in 1 ml of dry THF. The obtained mixture is stirred for 30 minutes, cooled to -78 ° and quenched with excess dry CO2. The obtained mixture is allowed to warm to RT, stirred overnight, diluted with CH 2 Cl 2, quenched with 2 ml 6 N HCl and the obtained mixture is refluxed for 1 hour. The obtained organic phase is washed with 1 N HCl, dried, and the obtained solvent mixture is evaporated. The evaporation residue is subjected to RP-HPLC. 1- (4-tert-Butyl-benzenesulfonyl) -1H-indole-2-carboxylic acid is obtained.

EXAMPLE 12:

1- (4-Morpholin-4-yl-benzenesulfonyl) -1Hindol-6-carboxylic acid methyl ester (compound of example 12 in Table 1)

A) 1- (4-BROMO-BENZENOSULPHONYL) 1H-INDOL-6-CARBOXYLIC ACID METHYL ESTER

To a solution of 500 mg of 1 H-indole acid methyl ester

6-Carboxylic acid in 15 ml DMF, 171 mg NaH (60% in mineral oil) are added. The obtained mixture is stirred for 15 minutes, treated with 789 mg of 4-bromo-benzenesulfonyl chloride, the obtained mixture is stirred overnight at RT and diluted with EtOAc. The obtained organic phase is washed with dried H2O1, and from the obtained solvent mixture is evaporated. The evaporation residue obtained is subjected to chromatography. 1- (4-Bromo-benzenesulfonyl) -1H-indole-6-carboxylic acid methyl ester is obtained.

B) 1- (4-MORFOLIN-4-IL-BENZENOSULPHONYL) -1H-INDOL-6-CARBOXYLIC ACID METHYL ESTER

To a solution of 30 mg of 1- (4-bromo-benzenesulfonyl) -1H-indole-6-carboxylic acid methyl ester in 2 ml of dry dioxane is added 8 mg of rac-BINAP, 81.8 mg (0.374 mmol) K3PO4 and 27 μΙ morpholine. The obtained mixture is purged with argon (30 minutes) and 3.58 mg of Pd (dba) 2 is added. The obtained mixture is stirred overnight at RT and diluted with EtOAc. The obtained organic phase is washed with H 2 O, each, and from the obtained solvent mixture is evaporated. The evaporation residue obtained is subjected to RP-HPLC. 1- (4-Morpholin-4-ylbenzenesulfonyl) -1H-indol-6-carboxylic acid methyl ester is obtained.

EXAMPLE 13:

1- (4-Morpholin-4-yl-benzenesulfonyl) -1 H -indol-6-carboxylic acid (compound of example 13 in Table 1)

To a solution of 120 mg of 1- (4-morpholin-4-yl-benzenesulfonyl) -1H-indol-6-carboxylic acid methyl ester in 40 ml of THF, a solution of 62.8 mg of LiOHxH2O in 4 ml of H2O is added. The obtained mixture is stirred for 18 hours at RT and the obtained mixture is diluted with 15 EtOAc. The obtained organic phase is washed with H2O, dried and the obtained solvent mixture is evaporated. The evaporation residue is subjected to RPHPLC. 1- (4-Morpholin-4-yl-benzenesulfonyl) -1H-indol-6-carboxylic acid is obtained.

EXAMPLE 14:

1- (4-tert-Butyl-benzenesulfonyl) -5-chloro-1H acid amide

indole-2-carboxylic acid (compound of example 14 in TABLE 1)

To a solution of 120 mg of 1- (4-tert-butyl-benzenesulfonyl) -5-chloro-1H-indol-2-carboxylic acid and 165 mg of NH4 Cl in 1 ml DMF, 1 ml diisopropyl ethylamine and 1 ml of PPA (50% in DMF) and 2 mg of DMAP 25 are added. The mixture is stirred for 18 h at room temperature. The obtained organic phase is filtered and the filtrate is subjected to RP-HPLC. 1- (4-tert-Butyl-benzenesulfonyl) -5-chloro-1H-indol-2-carboxylic acid amide is obtained.

Similarly as described in Example 14, but using the appropriate starting materials, the compounds of Examples 15 to 22 as shown and characterized in Table 1 are obtained. EXAMPLE 23:

1- (4-tert-Butyl-benzenesulfonyl) -5-chloro-1H-indol-2-carbonitrile (compound of example 23I in TABLE 1)

To a solution of 440 mg of 1- (4-tert-butyl5-benzenesulfonyl) -5-chloro-1H-indole-2-carboxylic acid amide in 6 ml of pyridine, 208 μΙ of POCI3 is added dropwise at 0 ° C. under argon atmosphere. The obtained mixture is stirred for 1 hour, solvent is evaporated and the obtained residue is diluted with EtOAc. The obtained mixture is washed with 1N HCl, sat. NaHCO3 and brine, and dried. The organic phase is concentrated and the obtained residue is chromatographed on silica gel. 1- (4-tert-Butyl-benzenesulfonyl) 5-chloro-1H-indol-2-carbonitrile is obtained.

Similarly as described in Example 23, but using the appropriate starting materials, the compounds of Examples 24 and 25 as shown and characterized in Table 1 are obtained.

EXAMPLES 26 AND 27:

1- (4-tert-Butyl-benzenesulfonyl) -1H-indole-6-carboxylic acid and 4-tert-butyl-N- [1- (4-tert-butyl-benzenesulfonyl) -1 H -indole-6 amide -carbonyl] -benzenesulfonamide (example compounds 26 and 27 in Table 1)

A solution of 140 mg of 1 H-indole-6-carboxylic acid amide and 23.8 mg of 18-crown-6 in 2 ml of anhydrous THF is cooled to 0 ° C under argon atmosphere. To the obtained mixture 1.1 ml of a solution of potassium tert-butoxide (1 M in THF) is added dropwise at 0 ° C, then a solution of 224 mg 4-tert-butylbenzenesulfonyl chloride in 4 ml of THF anhydrous. The obtained mixture is stirred for 1 hour at this temperature and the solvent is evaporated. The obtained residue is diluted with EtOAc. The obtained mixture is washed with brine and dried. The obtained organic phase is concentrated and the obtained residue is chromatographed on RP18.

1- (4-tert-Butyl-benzenesulfonyl) -1H-indole-6-carboxylic acid amide and 4-tert-butyl-N- [1- (4-tert-Butyl-benzenesulfonyl) -1 H -indole-6 -carbonyl] -benzenesulfonamide are obtained.

The compounds as described in Examples 1 to 27 above are compounds of the formula ^ 2ex jj ^ 1 Εχ Iex

THE

wherein R-iex and R2ex are as defined in Table 1 below and obtained analogously to the methods as described above. Melting points of the corresponding compounds obtained are also set forth in Table 1.

Example R-IEX R2EX M.p. (0C) 1 OR when given MH + or MNa + 1 -O3C / 198 to 202 J1 I)> -COOH 2 -O3C 115a 120 3 -O3C 135 to 136 4 -CfcC \ 250 to 252 HOOC 5 COOH 420 MH + 6 -O3C O 391 MH + i) OOX | 7/204 to 206 [fYN> COOH 8 -Ot ;: 403 MH + c1 / 4H2 CO2 Example s.-n-1 Riex R2EX M.p. (0C) 1 OR when given MH + or MNa +. 9 COOH 368 MH + 10/443 MNa + | I)> -COOH 11 -Cfc N 60 to 63 HOOC12 -OCo \ iCOOCH, 65 to 70 13 OO V ^^^ COOH 268 to 270 14 -CfcC \ 88 to 90 NH ^ 15 -CfcC \ n 460 MH + O ^ N ^^ The 16 -CfcC! 476 MH + O O j. Example 1 R1EX R2EX M.p. (0C) 1 OR when given MH + or MNa + 18 -cfcc \ 404 MH + CH3n H - \\ O 19 -CfcC \ 441 MNa + CH3n N- / CH3 / O 20 -CfcC \ n 481 MNa + O-C1 · 21 -CfcC \ 390 MH + NH 2 - \ O 22 -CfcC \ 432 MH + ch3 Vn-a / h \\ CH3 / xO 23 -CfcC 168 to 170 24 -CfcC \ 163 to 165 NC-I 25 -CfcC \ 164 to 165 CN Example RiEX R2EX Pf (0C) 1 OR when given MH + or MNa + 26 -CfcC O 223 to 225 / ^ j · ^ Jjp NH2 TI CH3 134 to 136 \ 1 nXJ ^ "·" · ζγγ ^ ο

Claims (9)

1. Use of a compound of the formula wherein R1 is (C1-12) alkyl, (C1-12) alkoxy, (C1-12) cycloalkyl optionally fused to heterocyclyl (C5-12) cycloalkenyl, optionally fused to heterocyclyl, (C6-18) aryl, optionally fused to heterocyclyl, heterocyclyl, halogen, halo (C1-4) alkyl, preferably CF3, (C1-4) alkoxy, halo (C - (4) alkoxy, (C1-4) alkylthio, wherein heterocyclyl is aliphatic or aromatic heterocyclyl comprising 3 to 12 ring members and 1 to 4 heteroatoms selected from N, O, S, optionally ringed with another ring system, wherein cycloalkyl, cycloalkenyl, aryl or heterocyclyl are unsubstituted or substituted by one or more halogen, halo (C1-4) alkyl, halo (C1-4) alkoxy, (C1-4) alkyl, (C1-4) alkoxy or (C1-4) alkylthio, R2 and R3 are different from each other and independently are hydrogen, hydroxycarbonyl, alkoxycarbonyl, e.g. (C1) alkoxycarbonyl, cyano, aminocarbonyl, where amino is NH 2, once or twice substituted amino or a cyclic amine, halogen, halo (C 1-4) alkyl, such as CF 3, (C 1-4) alkyl, (C 1-4) alkoxy, (C 1-4) Alkylthio in free or salt form in the manufacture of a medicament for the treatment of disorders that are mediated by CCR9 activity, preferably inflammatory bowel disease. Use according to claim 1, wherein in a compound of the formula (I) R 1 is (C-M2) alkyl, for example tert-butyl, (C1-12) alkoxy, (C3.12) cycloalkyl, optionally fused to heterocyclyl, (C5-12) cycloalkenyl, optionally fused to heterocyclyl, (C6-8) aryl, optionally fused to heterocyclyl, heterocyclyl, halogen, e.g. chlorine, halo (C1-4) alkyl, preferably CF3, (C1-4) alkoxy, halo (C1-4) alkoxy, (C1.4) alkylthio, wherein heterocyclyl is aliphatic or aromatic heterocyclyl comprising 3 to 12 ring members and 1 to 4 heteroatoms selected from N, O, S, optionally ringed with another ring system, for example 4-morpholin-4-yl or 4-oxazol-5-yl, wherein cycloalkyl, cycloalkenyl, aryl or heterocyclyl are unsubstituted or substituted by one or more halogen, halo (C1 .4) alkyl, halo (C1-4) alkoxy, (C1-4) alkyl, (C1-4) alkoxy or (C1-4) alkyl, R2 and R3 are different from each other and independently are hydrogen, hydroxycarbonyl, alkoxycarbonyl, for example (C1.4) alkoxycarbonyl, such as, for example, methoxycarbonyl, cyano, aminocarbonyl, where amino is NH2, one or two-substituted amino or a cyclic amine, for example amino, methylamino, dimethylamino, isopropylamino, piperidin-1-yl, morpholin-4-yl, 4-methyl-piperazin-1-yl, halogen, (C1-4) alkyl halo such as CF3, (C1-4) alkyl, (C1- alkoxy, (C1-4) alkylthio. 3. A compound of the formula wherein R 1 is branched (C 4-12) alkyl, preferably tert-butyl, (C 1-12) alkoxy, (C 3-12) cycloalkyl, optionally fused to heterocyclyl, (C5-12) cycloalkenyl, optionally fused to heterocyclyl, (C6-12), optionally fused to heterocyclyl, heterocyclyl, chlorine or fluorine, (C1-4) alkyl halo, preferably CF3, (C1-4 alkoxy, halo (C1-4) alkoxy, (C1.4) alkylthio, wherein heterocyclyl is aliphatic or aromatic heterocyclyl comprising 3 to 12 ring members and 1 to 4 heteroatoms selected from N, O, S, optionally ringed with another ring system, wherein cycloalkyl, cycloalkenyl, aryl or heterocyclyl are unsubstituted or substituted by one or more halogen, halo (C1-4) alkyl, halo (C1-4) alkoxy, (C1-4) alkyl, (C1-4) alkoxy or (C1-4) alkylthio, R2 and R3 are different from each other and independently are hydrogen, hydroxycarbonyl, alkoxycarbonyl, e.g. (C1-4) alkoxycarbonyl, cyano, aminocarbonyl, wherein amino is NH2, amino substituted once or twice or a cyclic amine, with the proviso that if R2 is 4-methylpiperazin-1-ylcarbonyl, then R2 is at position 3 of the ring system, indole-7-carboxylic acid 1- (4-tert-butyl-benzenesulfonyl) -1 H -indole-3-carboxylic acid, chlorine, halo (C 1-4) alkyl such as CF3, (C1-4) alkyl, (C1-4) alkoxy (C1-4) alkylthio with the proviso that a compound of the formula <formula> formula see original document page 71 </formula> and a compound of the formula <formula> formula see original document page 71 </formula> are deleted. 4. A compound selected from the group consisting of 1- (4-tert-butyl-benzenesulfonyl) -5-chloro-1 H -indol-2-carboxylic acid, 1- (4-tert-butyl-benzenesulfonyl) -1 H -indol-6-carboxylic acid 1- (4-tert-butyl-benzenesulfonyl) -1-methyl acid ester 5-chloro-1- (4-morpholin-4-yl) -benzenesulfonyl) -1H-indol-3 - carboxylic acid 1- (4-tert-butyl-benzenesulfonyl) -5-chloro-1H-indol-3-carboxylic acid, 5-chloro-1- (4-oxazol-5-yl) -benzenesulfonyl) -1 H-indole-3-carboxylic acid, 5-chloro-1- (4-trifluoromethyl-benzenesulfonyl) -1 H-indol-3-carboxylic acid, 5-chloro-1- (4-chloro-benzenesulfonyl) -1 H- indol-3-carboxylic acid 5-chloro-1- (4-morpholin-4-yl-benzenesulfonyl) -1H-indol-2-carboxylic acid 1- (4-tert-butyl-benzenesulfonyl) -1 H- indol-2-carboxylic acid 1- (4-morpholin-4-yl-benzenesulfonyl) -1Hindol-6-carboxylic acid methyl ester 1- (4-morpholin-4-yl-benzenesulfonyl) -1H-indol -6-carboxylic acid 1- (4-tert-butyl-benzenesulfonyl) -amide 5-chloro-1H-indol2-carboxylic acid, [1- (4-tert-butyl-benzenesulfonyl) -5-chloro-1H-indol-3-yl] -morpholin-4-yl-methanone, 2 - {[ 1- (4-tert-Butyl-benzenesulfonyl) -5-chloro-1H-indol-3-carbonyl] amino} -2-methyl-propionic, [1- (4-tert-Butyl-benzenesulfonyl) -5-chloro -1 H-indol-3-yl] - (4-methylpiperazin-1-yl) methanone, 1- (4-tert-butyl-benzenesulfonyl) -5-chloro-1 Hindol-3-carboxylic acid methylamide, dimethylamide 1- (4-tert-Butyl-benzenesulfonyl) -5-chloro-1 H -indol-3-carboxylic acid, [1- (4-tert-Butyl-benzenesulfonyl) -5-chloro-1H-indol-3-yl] 1- (4-tert-Butyl-benzenesulfonyl) -5-chloro-1H-indol-3-carboxylic acid amide, 1- (4-tert-butyl-benzenesulfonyl) -5-chloro-acid-piperidin-1-methanone . 1H-indol-3-carboxylic, 1- (4-tert-butyl-benzenesulfonyl) -1H-indol-6-carbonitrile, 1- (4-tert-butyl-benzenesulfonyl) -5-chloro-1H-indol2-carbonitrile, 1 - (4-tert-Butyl-benzenesulfonyl) -1H-indol-3-carbonitrile, 1- (4-tert-Butyl-benzenesulfonyl) -1H-indol-6-carboxylic acid amide and 4-tert-butyl-N- [1- (4-tert-Butyl-benzenesulfonyl) -1H-indol-6-carbonyl] -benzenesulfonamide, A compound according to claim 3 or 4, in the form of a salt. A compound according to any one of claims 3 to 5 for use as a pharmaceutical product. A pharmaceutical composition comprising a compound as defined in any one of claims 3 to 5, in association with at least one pharmaceutically acceptable excipient. A pharmaceutical combination comprising a compound as defined in any one of claims 3 to 5, or a pharmaceutical composition as defined in claim 7, and further comprising a second drug substance. A method for treating disorders that are mediated by CCR9 activity, the treatment of which comprises administering to a subject in need of such treatment a therapeutically effective amount of a compound as defined in any one of claims 3 to 5, or a pharmaceutical composition as defined in claim 6, optionally in combination with a second drug substance.